LOCUS       Exported                8082 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000542 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8082)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     This file was created using tools provided by Twist Bioscience
FEATURES             Location/Qualifiers
     source          1..8082
                     /organism="BBF10K_000542 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..3588
                     /codon_start=1
                     /product="ITGAE integrin subunit alpha E"
                     /label=CD103
                     /translation="MWLFHTLLCIASLALLAAFNVDVARPWLTPKGGAPFVLSSLLHQD
                     PSTNQTWLLVTSPRTKRTPGPLHRCSLVQDEILCHPVEHVPIPKGRHRGVTVVRSHHGV
                     LICIQVLVRRPHSLSSELTGTCSLLGPDLRPQAQANFFDLENLLDPDARVDTGDCYSNK
                     EGGGEDDVNTARQRRALEKEEEEDKEEEEDEEEEEAGTEIAIILDGSGSIDPPDFQRAK
                     DFISNMMRNFYEKCFECNFALVQYGGVIQTEFDLRDSQDVMASLARVQNITQVGSVTKT
                     ASAMQHVLDSIFTSSHGSRRKASKVMVVLTDGGIFEDPLNLTTVINSPKMQGVERFAIG
                     VGEEFKSARTARELNLIASDPDETHAFKVTNYMALDGLLSKLRYNIISMEGTVGDALHY
                     QLAQIGFSAQILDERQVLLGAVGAFDWSGGALLYDTRSRRGRFLNQTAAAAADAEAAQY
                     SYLGYAVAVLHKTCSLSYIAGAPRYKHHGAVFELQKEGREASFLPVLEGEQMGSYFGSE
                     LCPVDIDMDGSTDFLLVAAPFYHVHGEEGRVYVYRLSEQDGSFSLARILSGHPGFTNAR
                     FGFAMAAMGDLSQDKLTDVAIGAPLEGFGADDGASFGSVYIYNGHWDGLSASPSQRIRA
                     STVAPGLQYFGMSMAGGFDISGDGLADITVGTLGQAVVFRSRPVVRLKVSMAFTPSALP
                     IGFNGVVNVRLCFEISSVTTASESGLREALLNFTLDVDVGKQRRRLQCSDVRSCLGCLR
                     EWSSGSQLCEDLLLMPTEGELCEEDCFSNASVKVSYQLQTPEGQTDHPQPILDRYTEPF
                     AIFQLPYEKACKNKLFCVAELQLATTVSQQELVVGLTKELTLNINLTNSGEDSYMTSMA
                     LNYPRNLQLKRMQKPPSPNIQCDDPQPVASVLIMNCRIGHPVLKRSSAHVSVVWQLEEN
                     AFPNRTADITVTVTNSNERRSLANETHTLQFRHGFVAVLSKPSIMYVNTGQGLSHHKEF
                     LFHVHGENLFGAEYQLQICVPTKLRGLQVVAVKKLTRTQASTVCTWSQERACAYSSVQH
                     VEEWHSVSCVIASDKENVTVAAEISWDHSEELLKDVTELQILGEISFNKSLYEGLNAEN
                     HRTKITVVFLKDEKYHSLPIIIKGSVGGLLVLIVILVILFKCGFFKRKYQQLNLESIRK
                     AQLKSENLLEEEN"
     primer_bind     complement(3589..3613)
                     /label=Gibson-rev
     terminator      3732..3779
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3784..3800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3959..4209
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(4212..4967)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      5058..5277
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    5697..5718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5733..5763
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5771..5787
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5795..5811
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5884..6729
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    6837..6988
                     /label=nnpp promoters
     CDS             7028..7843
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      complement(7953..7996)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     8014..8030
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      8031..8062
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgtggctg
       61 tttcatacat tgttgtgtat tgcatccctc gcattgttgg ctgcatttaa tgtcgatgtt
      121 gctcggccat ggctcactcc aaaaggtggg gcaccgtttg tcctttccag tttgcttcat
      181 caagatcctt caactaatca aacgtggctt ctcgtcactt ctcctaggac gaaacggacg
      241 ccaggaccac ttcatcgatg ttcacttgta caagatgaaa ttctttgtca tcctgtcgaa
      301 catgtcccta ttccaaaagg gcggcatcgc ggagttacag ttgtcaggtc ccatcatggg
      361 gtcctcatat gtatacaagt tttggttagg cggccacatt cattgagttc tgaacttacg
      421 ggtacttgtt cattgctcgg accagatttg cgaccgcaag ctcaagcaaa tttctttgat
      481 ttggaaaatc ttctcgatcc tgatgcaagg gtagatacgg gtgattgtta ttcaaataaa
      541 gaaggtggtg gagaagatga tgtaaataca gcacggcaaa ggcgtgcact tgaaaaggaa
      601 gaagaagaag ataaagaaga agaagaagat gaagaagaag aagaagcagg gacggaaatt
      661 gctattattt tggatggttc agggtccatt gacccaccag attttcaacg cgctaaagac
      721 tttatttcca atatgatgcg caatttctat gagaaatgtt ttgaatgtaa ttttgcattg
      781 gtacaatatg gcggggttat tcaaacggaa ttcgatctcc gggattctca agatgtcatg
      841 gcgagtctcg ctcgagtcca aaatattacg caagtcggga gtgttacaaa gactgctagt
      901 gctatgcaac atgttctcga tagtatattt acatcctctc atggatctag gcgtaaagcg
      961 tccaaagtca tggtagttct cacagatggc ggtatatttg aagatccact taatcttact
     1021 actgtcatta atagtcccaa aatgcaagga gttgaacggt ttgcaattgg tgtcggggaa
     1081 gaatttaaat ccgcacgcac ggctagggaa cttaatttga ttgcttcaga tccggatgaa
     1141 acacatgcat ttaaagtcac taattatatg gctcttgatg gacttctctc taaactccgc
     1201 tataatatta tttctatgga agggacagtc ggggatgcat tgcattatca acttgcgcaa
     1261 ataggtttct ctgcacaaat tctcgatgaa cggcaagtcc ttctcggagc agtcggtgct
     1321 tttgattgga gtggcggggc tcttttgtat gatacacggt ctcgccgggg ccgctttctc
     1381 aatcaaactg ctgcggcagc tgctgatgca gaagcagcgc aatatagtta tctcggttat
     1441 gcagtcgcag ttctccataa aacatgttca ttgtcctata ttgcaggggc tccgaggtat
     1501 aaacatcatg gggctgtttt cgaattgcaa aaggaaggtc gcgaagcatc ctttcttcct
     1561 gttctcgaag gagaacaaat gggttcctat tttggttctg aactctgtcc agtagatata
     1621 gatatggatg gatcaacaga tttcttgctc gttgcagctc ctttctatca tgtccatggg
     1681 gaagaagggc gtgtttatgt ctatcgcctt tccgaacaag atggaagctt ttctctcgct
     1741 cgcatactct ccggtcatcc agggtttact aatgcaaggt ttgggtttgc gatggctgcg
     1801 atgggtgatc tttcccaaga taaactcact gatgttgcaa ttggtgctcc attggaagga
     1861 tttggtgctg atgatggagc ttcctttgga tcagtttata tttataatgg gcattgggat
     1921 ggactcagtg cgtctcctag tcaacggatt cgcgcgtcta ctgttgctcc tggactccaa
     1981 tattttggta tgtctatggc aggtgggttt gatatttccg gagatggttt ggctgatatt
     2041 acggtaggta ctctcggaca agctgtcgtc tttcggtcac gaccagtagt ccgccttaaa
     2101 gtctccatgg catttacacc atctgcattg cctatagggt ttaatggtgt cgttaatgtc
     2161 aggctctgtt ttgaaataag ttccgttact acggctagtg aatctgggct ccgggaagct
     2221 ctcttgaatt ttactctcga tgtagatgtt gggaaacaaa ggcgccgact ccaatgtagt
     2281 gatgttcgct cttgtttggg gtgtctcagg gaatggtcaa gtggctccca attgtgtgaa
     2341 gatcttcttt tgatgcctac agaaggagaa ctctgtgaag aagattgctt ttcaaatgca
     2401 tcagtcaaag tttcttatca actccaaaca cctgaaggtc aaacagatca tcctcaacct
     2461 attcttgatc gctatacaga accgtttgca atctttcaat tgccgtatga gaaagcgtgt
     2521 aagaataaac tcttttgtgt tgctgaactc caattggcta caactgtcag tcaacaagaa
     2581 ttggttgtcg gtttgacaaa agaactcaca ttgaatatta atctcacgaa ttctggtgaa
     2641 gattcataca tgacatctat ggcgctcaat tatcctcgca atttgcaact caaacggatg
     2701 cagaaaccac cttctccaaa tattcaatgt gatgatcctc aaccagtcgc tagtgtattg
     2761 attatgaatt gtaggattgg gcatcctgtc cttaaacgat ccagtgcgca tgtatcagtc
     2821 gtttggcaac ttgaagaaaa tgcttttcct aatcgaactg cagatataac agtaacagtc
     2881 actaattcta atgaacgccg gtcccttgca aatgaaacac atactttgca atttcgccat
     2941 ggatttgtag cagtcctttc caaaccttct attatgtatg taaataccgg acaaggtctc
     3001 tcccatcata aagaatttct cttccatgtt catggtgaaa atttgtttgg agcagagtat
     3061 caattgcaaa tttgtgtacc aacaaaactc cgtggacttc aagtagtcgc tgtcaagaaa
     3121 ctcacacgga cacaagcaag tacggtttgt acatggagtc aagaacgcgc gtgtgcgtat
     3181 tcctcagtcc aacatgttga agaatggcat tctgtaagtt gtgtcattgc ttctgataaa
     3241 gaaaatgtta cagttgctgc tgaaattagt tgggatcatt ctgaagaact cctgaaagat
     3301 gttacagaat tgcaaattct cggtgaaata tcctttaata aatccctcta tgaaggactc
     3361 aatgctgaaa atcatcgcac gaagattacg gtcgtctttc tcaaagatga aaagtatcat
     3421 agtctcccaa ttataattaa agggtcagtc ggcggactcc tcgtattgat agtcattttg
     3481 gtgattctct tcaaatgtgg tttctttaaa aggaaatatc aacaattgaa ccttgaatcc
     3541 attcggaaag cgcaactcaa aagtgaaaat ttgctcgaag aggagaacgg atccgccacc
     3601 aacttcagcc tgctatacgt ggctagcatg agacatcgtg ttgcacgcat aggtgtaggg
     3661 ataacagggt aatctctcca tcctcgcaag aacccgctga ggtgtcaatc gtcggagccg
     3721 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     3781 atggtcatag ctgtttcctg agagcttggc aggtgtcgcg tatgcatcga tcacgtgtcc
     3841 acgttcttta atagtggact cttgttccaa actggaacaa cactcggatc gatccggcgc
     3901 gcaccgtggg aaaaactcca ggtagaggta cacacgcgga tagccaattc agagtaataa
     3961 actgtgataa tcaaccctca tcaatgatga cgaactaacc cccgatatca ggtcacatga
     4021 cgaagggaaa gagaaggaaa tcaactgtga caaactgccc tcaaatttgg cttccttaaa
     4081 aattacagtt caaaaagtat gagaaaatcc atgcaggctg aaggaaacag caaaactgtg
     4141 acaaattacc ctcagtaggt cagaacaaat gtgacgaacc accctcaaat ctgtgacaga
     4201 taaccctcag actatcctgt cgtcatggaa gtgatatcgc ggaaggaaaa tacgatatga
     4261 gtcgtctggc ggcctttctt tttctcaatg tatgagaggc gcattggagt tctgctgttg
     4321 atctcattaa cacagacttg caggaagcgg cggcggaagt caggcatacg ctggtaactt
     4381 tgaggcagct ggtaacgctc tatgatccag tcgattttca gagacacgat gcctgagcca
     4441 tccggcttac gatactgaca cagggattcg tataaacgca tggcatacgg attggtgatt
     4501 tcttttgttt cactaagccg aaactgcgta aaccggttct gtaacccgat aaagaaggga
     4561 atgagatatg ggttgatatg tacactgtaa agccctctgg atggactgtg cgcacgtttg
     4621 ataaaccaag gaaaagattc atagcctttt tcatcaccgg catcctcttc agggcgataa
     4681 aaaaccactt ccttccccgc gaaactcttc aatgcctgcc gtatatcctt actggcttcc
     4741 gcagaggtca atccgaatat ttcagcatat ttagcaacat ggatctcgca gataccgtca
     4801 tgttcctgta gggtgccatc agattttctg atctggtcaa cgaacagata cagcatacgt
     4861 ttttgatccc gggagagact atatgccgcc tcagtgaggt cgtttgactg gacgattcgc
     4921 gggctatttt tacgtttctt gtgattgata accgctgttt ccgccatgac agatccatgt
     4981 gaagtgtgac aagtttttag attgtcacac taaataaaaa agagtcaata agcagggata
     5041 actttgtgaa aaaacagctt cttctgaggg caatttgtca cagggttaag ggcaatttgt
     5101 cacagacagg actgtcattt gagggtgatt tgtcacactg aaagggcaat ttgtcacaac
     5161 accttctgta gaaccagcat ggataaaggc ctacaaggcg ctctaaaaaa gaagatctaa
     5221 aaactataaa aaaaataatt ataaaaatat ccccgtggat aagtggataa ccccaaggga
     5281 agttttttca ggcatcgtgt gtaagcagaa tatataagtg ctgttccctg gtgcttcctc
     5341 gctcactcga aattcccggg gatagcttta tgcttgtaaa ccgttttgtg aaaaaatttt
     5401 taaaataaaa aaggggacct ctagggtccc caattaatta gtaatataat ctattaaagg
     5461 tcattcaaaa ggtcatccac cggatcaatt cccctgctcg cgcaggctgg gtgccaagct
     5521 ctcgggtaac atcaaggccc gatccttgga gcccttgccc tcccgcacga tgatcgtgcc
     5581 gtgatcgaaa tccagatcct tgacccgcag ttgcaaaccc tcactgatcc gattcattaa
     5641 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5701 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5761 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5821 gccaagcgcg gtacctgtcg cggcaacgcg ctaacagacg tagtaagaac caccagcatt
     5881 gtaatgctgg ctaaagtcac tttcctgagc tgtataacga tgagcgattt tactttttct
     5941 ggctatgaat tggcctgctt tgtaacacac tccggtctat cccgtagcgc cgggcatatc
     6001 ctgtcgcaat gtgcaaatct cgcggcaaca accagtgaat acttcattca caagcctcac
     6061 cgcctgatcg cggcagaaac tggttatagc caatcaaccg tcgttcgtgc attccgtgaa
     6121 gctgtaaaca aaggtatcct gtctgtagag attgttatcg gcgatcaccg tgaacgtcgc
     6181 gctaacctgt accggtttac accatccttt ttggccttcg cacaacaagc caaaaatgcg
     6241 ctgatagaaa gcaaattaaa gatctcttca gcggcaacca aggttaaagc tgttctcgct
     6301 aagacattgg ctttatttaa ttttttatcc acacccccat gtcaaaatga taccccctcc
     6361 ccctgtcagg atgacgtggc aataaagaat aagaagtcac aagttaaaaa aacaaaaaga
     6421 tcagtttccg gcggtgccgg aacaaccagc ctcaaaaaat tgacttcatg gatcgctaag
     6481 gcaaaagcaa aggctgacaa tctgcggtta tccaaaaaac gcactcaaaa acatgagttc
     6541 aagcagaaag tagaggcggc tgcgcggaaa tatgcttacc tgaagaacaa gcgttcgcct
     6601 gatattggcg ggatatcaaa cttcgataac ctaccgcatt gcatgacggt aaacgaagct
     6661 cttaatgcgg ttttagccaa aaataaagat aacgaacaat ggggtatacc ggcaggattc
     6721 agagggtaat gaattgctct aattataacc atgcatactt tcaacacctc tagtttgcca
     6781 tgaggcaaac tcataggtgt cctggtaaga ggacactgtt gccaaaactg gacgccccat
     6841 tattgcaatt aataaacaac taacggacaa ttctacctaa caataagtgg agttgcggcc
     6901 ggccgcgatc cgctagcaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     6961 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     7021 gggtgttatg agccatattc aacgggaaac gtcttgctcg cgcccgcgat taaattccaa
     7081 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     7141 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     7201 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     7261 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     7321 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     7381 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     7441 ttgtcctttt aacagcgatc gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa
     7501 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     7561 ctggaaagaa atgcataaac tgttgccatt ctcaccggat tcagtcgtca ctcatggtga
     7621 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     7681 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     7741 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     7801 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     7861 gttgtaacac tggcgagctc tgccacctga cgtctaagaa accattatta tcatgacatt
     7921 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
     7981 cgctaaggat gatttctgca cctgcaccag tcagtaaaac gacggccagt agtcaaaagc
     8041 ctccgaccgg aggcttttga cttggttcag gtggagtggg ag
//
