LOCUS       Exported                5172 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000536 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5172)
  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..5172
                     /organism="BBF10K_000536 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..678
                     /codon_start=1
                     /product="regulator of G protein signaling 1"
                     /label=RGS1
                     /translation="MRAAAISTPKLDKMPGMFFSANPKELKGTTHSLLDDKMQKRRPKT
                     FGMDMKAYLRSMIPHLESGMKSSKSKDVLSAAEVMQWSQSLEKLLANQTGQNVFGSFLK
                     SEFSEENIEFWLACEDYKKTESDLLPCKAEEIYKAFVHSDAAKQINIDFRTRESTAKKI
                     KAPTPTCFDEAQKVIYTLMEKDSYPRFLKSDIYLNLLNDLQANSLK"
     primer_bind     complement(679..703)
                     /label=Gibson-rev
     terminator      822..869
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(874..890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1049..1299
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1302..2057)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2148..2367
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2787..2808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2823..2853
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2861..2877
                     /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     2885..2901
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2974..3819
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    3927..4078
                     /label=nnpp promoters
     CDS             4118..4933
                     /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(5043..5086)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5104..5120
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5121..5152
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgagagcc
       61 gccgccatca gcacccccaa gctggacaag atgcccggca tgttcttcag cgccaacccc
      121 aaggagctga agggcaccac ccacagcctg ctggacgaca agatgcagaa gagaagaccc
      181 aagaccttcg gcatggacat gaaggcctac ctgagaagca tgatccccca cctggagagc
      241 ggcatgaaga gcagcaagag caaggacgtg ctgagcgccg ccgaggtgat gcagtggagc
      301 cagagcctgg agaagctgct ggccaaccag accggccaga acgtgttcgg cagcttcctg
      361 aagagcgagt tcagcgagga gaacatcgag ttctggctgg cctgcgagga ctacaagaag
      421 accgagagcg acctgctgcc ctgcaaggcc gaggagatct acaaggcctt cgtgcacagc
      481 gacgccgcca agcagatcaa catcgacttc agaaccagag agagcaccgc caagaagatc
      541 aaggccccca cccccacctg cttcgacgag gcccagaagg tgatctacac cctgatggag
      601 aaggacagct accccagatt cctgaagagc gacatctacc tgaacctgct gaacgacctg
      661 caggccaaca gcctgaaggg atccgccacc aacttcagcc tgctagtgaa ctgaccgaat
      721 cctcgtttcg tcgaagctcg aaaggtaggg ataacagggt aataggacct tctaatccac
      781 ccctcgctga ggtgtcaatc gtcggagccg ctgagcaata actagcataa ccccttgggg
      841 cctctaaacg ggtcttgagg ggttttttgc atggtcatag ctgtttcctg agagcttggc
      901 aggtgtcgcg tatgcatcga tcacgtgtcc acgttcttta atagtggact cttgttccaa
      961 actggaacaa cactcggatc gatccggcgc gcaccgtggg aaaaactcca ggtagaggta
     1021 cacacgcgga tagccaattc agagtaataa actgtgataa tcaaccctca tcaatgatga
     1081 cgaactaacc cccgatatca ggtcacatga cgaagggaaa gagaaggaaa tcaactgtga
     1141 caaactgccc tcaaatttgg cttccttaaa aattacagtt caaaaagtat gagaaaatcc
     1201 atgcaggctg aaggaaacag caaaactgtg acaaattacc ctcagtaggt cagaacaaat
     1261 gtgacgaacc accctcaaat ctgtgacaga taaccctcag actatcctgt cgtcatggaa
     1321 gtgatatcgc ggaaggaaaa tacgatatga gtcgtctggc ggcctttctt tttctcaatg
     1381 tatgagaggc gcattggagt tctgctgttg atctcattaa cacagacttg caggaagcgg
     1441 cggcggaagt caggcatacg ctggtaactt tgaggcagct ggtaacgctc tatgatccag
     1501 tcgattttca gagacacgat gcctgagcca tccggcttac gatactgaca cagggattcg
     1561 tataaacgca tggcatacgg attggtgatt tcttttgttt cactaagccg aaactgcgta
     1621 aaccggttct gtaacccgat aaagaaggga atgagatatg ggttgatatg tacactgtaa
     1681 agccctctgg atggactgtg cgcacgtttg ataaaccaag gaaaagattc atagcctttt
     1741 tcatcaccgg catcctcttc agggcgataa aaaaccactt ccttccccgc gaaactcttc
     1801 aatgcctgcc gtatatcctt actggcttcc gcagaggtca atccgaatat ttcagcatat
     1861 ttagcaacat ggatctcgca gataccgtca tgttcctgta gggtgccatc agattttctg
     1921 atctggtcaa cgaacagata cagcatacgt ttttgatccc gggagagact atatgccgcc
     1981 tcagtgaggt cgtttgactg gacgattcgc gggctatttt tacgtttctt gtgattgata
     2041 accgctgttt ccgccatgac agatccatgt gaagtgtgac aagtttttag attgtcacac
     2101 taaataaaaa agagtcaata agcagggata actttgtgaa aaaacagctt cttctgaggg
     2161 caatttgtca cagggttaag ggcaatttgt cacagacagg actgtcattt gagggtgatt
     2221 tgtcacactg aaagggcaat ttgtcacaac accttctgta gaaccagcat ggataaaggc
     2281 ctacaaggcg ctctaaaaaa gaagatctaa aaactataaa aaaaataatt ataaaaatat
     2341 ccccgtggat aagtggataa ccccaaggga agttttttca ggcatcgtgt gtaagcagaa
     2401 tatataagtg ctgttccctg gtgcttcctc gctcactcga aattcccggg gatagcttta
     2461 tgcttgtaaa ccgttttgtg aaaaaatttt taaaataaaa aaggggacct ctagggtccc
     2521 caattaatta gtaatataat ctattaaagg tcattcaaaa ggtcatccac cggatcaatt
     2581 cccctgctcg cgcaggctgg gtgccaagct ctcgggtaac atcaaggccc gatccttgga
     2641 gcccttgccc tcccgcacga tgatcgtgcc gtgatcgaaa tccagatcct tgacccgcag
     2701 ttgcaaaccc tcactgatcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2761 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2821 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2881 cacacaggaa acagctatga ccatgattac gccaagcgcg gtacctgtcg cggcaacgcg
     2941 ctaacagacg tagtaagaac caccagcatt gtaatgctgg ctaaagtcac tttcctgagc
     3001 tgtataacga tgagcgattt tactttttct ggctatgaat tggcctgctt tgtaacacac
     3061 tccggtctat cccgtagcgc cgggcatatc ctgtcgcaat gtgcaaatct cgcggcaaca
     3121 accagtgaat acttcattca caagcctcac cgcctgatcg cggcagaaac tggttatagc
     3181 caatcaaccg tcgttcgtgc attccgtgaa gctgtaaaca aaggtatcct gtctgtagag
     3241 attgttatcg gcgatcaccg tgaacgtcgc gctaacctgt accggtttac accatccttt
     3301 ttggccttcg cacaacaagc caaaaatgcg ctgatagaaa gcaaattaaa gatctcttca
     3361 gcggcaacca aggttaaagc tgttctcgct aagacattgg ctttatttaa ttttttatcc
     3421 acacccccat gtcaaaatga taccccctcc ccctgtcagg atgacgtggc aataaagaat
     3481 aagaagtcac aagttaaaaa aacaaaaaga tcagtttccg gcggtgccgg aacaaccagc
     3541 ctcaaaaaat tgacttcatg gatcgctaag gcaaaagcaa aggctgacaa tctgcggtta
     3601 tccaaaaaac gcactcaaaa acatgagttc aagcagaaag tagaggcggc tgcgcggaaa
     3661 tatgcttacc tgaagaacaa gcgttcgcct gatattggcg ggatatcaaa cttcgataac
     3721 ctaccgcatt gcatgacggt aaacgaagct cttaatgcgg ttttagccaa aaataaagat
     3781 aacgaacaat ggggtatacc ggcaggattc agagggtaat gaattgctct aattataacc
     3841 atgcatactt tcaacacctc tagtttgcca tgaggcaaac tcataggtgt cctggtaaga
     3901 ggacactgtt gccaaaactg gacgccccat tattgcaatt aataaacaac taacggacaa
     3961 ttctacctaa caataagtgg agttgcggcc ggccgcgatc cgctagcaaa gccacgttgt
     4021 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     4081 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     4141 gtcttgctcg cgcccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     4201 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     4261 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     4321 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     4381 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
     4441 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4501 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4561 cctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4621 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac tgttgccatt
     4681 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4741 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4801 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4861 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4921 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcgagctc tgccacctga
     4981 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca
     5041 gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgca cctgcaccag
     5101 tcagtaaaac gacggccagt agtcaaaagc ctccgaccgg aggcttttga cttggttcag
     5161 gtggagtggg ag
//
