LOCUS       Exported                5787 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000515 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  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..5787
                     /organism="BBF10K_000515 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1293
                     /codon_start=1
                     /product="HHLA2 HERV–H LTR-associating protein 2"
                     /label=B7H7
                     /translation="MKAQTALSFFLILITSLSGSQGIFPLAFFIYVPMNEQIVIGRLDE
                     DIILPSSFERGSEVVIHWKYQDSYKVHSYYKGSDHLESQDPRYANRTSLFYNEIQNGNA
                     SLFFRRVSLLDEGIYTCYVGTAIQVITNKVVLKVGVFLTPVMKYEKRNTNSFLICSVLS
                     VYPRPIITWKMDNTPISENNMEETGSLDSFSINSPLNITGSNSSYECTIENSLLKQTWT
                     GRWTMKDGLHKMQSEHVSLSCQPVNDYFSPNQDFKVTWSRMKSGTFSVLAYYLSSSQNT
                     IINESRFSWNKELINQSDFSMNLMDLNLSDSGEYLCNISSDEYTLLTIHTVHVEPSQET
                     ASHNKGLWILVPSAILAAFLLIWSVKCCRAQLEARRSRHPADGAQQERCCVPPGERCPS
                     APDNGEENVPLSGKV"
     primer_bind     complement(1294..1318)
                     /label=Gibson-rev
     terminator      1437..1484
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1489..1505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1664..1914
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1917..2672)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2763..2982
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3402..3423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3438..3468
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3476..3492
                     /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     3500..3516
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3589..4434
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4542..4693
                     /label=nnpp promoters
     CDS             4733..5548
                     /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(5658..5701)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5719..5735
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5736..5767
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgaaggcc
       61 cagaccgccc tgagcttctt cctgatcctg atcaccagcc tgagcggcag ccagggcatc
      121 ttccccctgg ccttcttcat ctacgtgccc atgaacgagc agatcgtgat cggcagactg
      181 gacgaggaca tcatcctgcc cagcagcttc gagagaggca gcgaggtggt gatccactgg
      241 aagtaccagg acagctacaa ggtgcacagc tactacaagg gcagcgacca cctggagagc
      301 caggacccca gatacgccaa cagaaccagc ctgttctaca acgagatcca gaacggcaac
      361 gccagcctgt tcttcagaag agtgagcctg ctggacgagg gcatctacac ctgctacgtg
      421 ggcaccgcca tccaggtgat caccaacaag gtggtgctga aggtgggcgt gttcctgacc
      481 cccgtgatga agtacgagaa gagaaacacc aacagcttcc tgatctgcag cgtgctgagc
      541 gtgtacccca gacccatcat cacctggaag atggacaaca cccccatcag cgagaacaac
      601 atggaggaga ccggcagcct ggacagcttc agcatcaaca gccccctgaa catcaccggc
      661 agcaacagca gctacgagtg caccatcgag aacagcctgc tgaagcagac ctggaccggc
      721 agatggacca tgaaggacgg cctgcacaag atgcagagcg agcacgtgag cctgagctgc
      781 cagcccgtga acgactactt cagccccaac caggacttca aggtgacctg gagcagaatg
      841 aagagcggca ccttcagcgt gctggcctac tacctgagca gcagccagaa caccatcatc
      901 aacgagagca gattcagctg gaacaaggag ctgatcaacc agagcgactt cagcatgaac
      961 ctgatggacc tgaacctgag cgacagcggc gagtacctgt gcaacatcag cagcgacgag
     1021 tacaccctgc tgaccatcca caccgtgcac gtggagccca gccaggagac cgccagccac
     1081 aacaagggcc tgtggatact ggtgcccagc gccatcctgg ccgccttcct gctgatctgg
     1141 agcgtgaagt gctgcagagc ccagctggag gccagaagaa gcagacaccc cgccgacggc
     1201 gcccagcagg agagatgctg cgtgcccccc ggcgagagat gccccagcgc ccccgacaac
     1261 ggcgaggaga acgtgcccct gagcggcaag gtgggatccg ccaccaactt cagcctgcta
     1321 gatagatgct ccgtcaagca ccagtgcata tccccattgt tagggataac agggtaatac
     1381 gtatggttct gtccaggtgc gctgaggtgt caatcgtcgg agccgctgag caataactag
     1441 cataacccct tggggcctct aaacgggtct tgaggggttt tttgcatggt catagctgtt
     1501 tcctgagagc ttggcaggtg tcgcgtatgc atcgatcacg tgtccacgtt ctttaatagt
     1561 ggactcttgt tccaaactgg aacaacactc ggatcgatcc ggcgcgcacc gtgggaaaaa
     1621 ctccaggtag aggtacacac gcggatagcc aattcagagt aataaactgt gataatcaac
     1681 cctcatcaat gatgacgaac taacccccga tatcaggtca catgacgaag ggaaagagaa
     1741 ggaaatcaac tgtgacaaac tgccctcaaa tttggcttcc ttaaaaatta cagttcaaaa
     1801 agtatgagaa aatccatgca ggctgaagga aacagcaaaa ctgtgacaaa ttaccctcag
     1861 taggtcagaa caaatgtgac gaaccaccct caaatctgtg acagataacc ctcagactat
     1921 cctgtcgtca tggaagtgat atcgcggaag gaaaatacga tatgagtcgt ctggcggcct
     1981 ttctttttct caatgtatga gaggcgcatt ggagttctgc tgttgatctc attaacacag
     2041 acttgcagga agcggcggcg gaagtcaggc atacgctggt aactttgagg cagctggtaa
     2101 cgctctatga tccagtcgat tttcagagac acgatgcctg agccatccgg cttacgatac
     2161 tgacacaggg attcgtataa acgcatggca tacggattgg tgatttcttt tgtttcacta
     2221 agccgaaact gcgtaaaccg gttctgtaac ccgataaaga agggaatgag atatgggttg
     2281 atatgtacac tgtaaagccc tctggatgga ctgtgcgcac gtttgataaa ccaaggaaaa
     2341 gattcatagc ctttttcatc accggcatcc tcttcagggc gataaaaaac cacttccttc
     2401 cccgcgaaac tcttcaatgc ctgccgtata tccttactgg cttccgcaga ggtcaatccg
     2461 aatatttcag catatttagc aacatggatc tcgcagatac cgtcatgttc ctgtagggtg
     2521 ccatcagatt ttctgatctg gtcaacgaac agatacagca tacgtttttg atcccgggag
     2581 agactatatg ccgcctcagt gaggtcgttt gactggacga ttcgcgggct atttttacgt
     2641 ttcttgtgat tgataaccgc tgtttccgcc atgacagatc catgtgaagt gtgacaagtt
     2701 tttagattgt cacactaaat aaaaaagagt caataagcag ggataacttt gtgaaaaaac
     2761 agcttcttct gagggcaatt tgtcacaggg ttaagggcaa tttgtcacag acaggactgt
     2821 catttgaggg tgatttgtca cactgaaagg gcaatttgtc acaacacctt ctgtagaacc
     2881 agcatggata aaggcctaca aggcgctcta aaaaagaaga tctaaaaact ataaaaaaaa
     2941 taattataaa aatatccccg tggataagtg gataacccca agggaagttt tttcaggcat
     3001 cgtgtgtaag cagaatatat aagtgctgtt ccctggtgct tcctcgctca ctcgaaattc
     3061 ccggggatag ctttatgctt gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg
     3121 gacctctagg gtccccaatt aattagtaat ataatctatt aaaggtcatt caaaaggtca
     3181 tccaccggat caattcccct gctcgcgcag gctgggtgcc aagctctcgg gtaacatcaa
     3241 ggcccgatcc ttggagccct tgccctcccg cacgatgatc gtgccgtgat cgaaatccag
     3301 atccttgacc cgcagttgca aaccctcact gatccgattc attaatgcag ctggcacgac
     3361 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     3421 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     3481 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcggtacc
     3541 tgtcgcggca acgcgctaac agacgtagta agaaccacca gcattgtaat gctggctaaa
     3601 gtcactttcc tgagctgtat aacgatgagc gattttactt tttctggcta tgaattggcc
     3661 tgctttgtaa cacactccgg tctatcccgt agcgccgggc atatcctgtc gcaatgtgca
     3721 aatctcgcgg caacaaccag tgaatacttc attcacaagc ctcaccgcct gatcgcggca
     3781 gaaactggtt atagccaatc aaccgtcgtt cgtgcattcc gtgaagctgt aaacaaaggt
     3841 atcctgtctg tagagattgt tatcggcgat caccgtgaac gtcgcgctaa cctgtaccgg
     3901 tttacaccat cctttttggc cttcgcacaa caagccaaaa atgcgctgat agaaagcaaa
     3961 ttaaagatct cttcagcggc aaccaaggtt aaagctgttc tcgctaagac attggcttta
     4021 tttaattttt tatccacacc cccatgtcaa aatgataccc cctccccctg tcaggatgac
     4081 gtggcaataa agaataagaa gtcacaagtt aaaaaaacaa aaagatcagt ttccggcggt
     4141 gccggaacaa ccagcctcaa aaaattgact tcatggatcg ctaaggcaaa agcaaaggct
     4201 gacaatctgc ggttatccaa aaaacgcact caaaaacatg agttcaagca gaaagtagag
     4261 gcggctgcgc ggaaatatgc ttacctgaag aacaagcgtt cgcctgatat tggcgggata
     4321 tcaaacttcg ataacctacc gcattgcatg acggtaaacg aagctcttaa tgcggtttta
     4381 gccaaaaata aagataacga acaatggggt ataccggcag gattcagagg gtaatgaatt
     4441 gctctaatta taaccatgca tactttcaac acctctagtt tgccatgagg caaactcata
     4501 ggtgtcctgg taagaggaca ctgttgccaa aactggacgc cccattattg caattaataa
     4561 acaactaacg gacaattcta cctaacaata agtggagttg cggccggccg cgatccgcta
     4621 gcaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     4681 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     4741 tattcaacgg gaaacgtctt gctcgcgccc gcgattaaat tccaacatgg atgctgattt
     4801 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     4861 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     4921 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     4981 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     5041 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     5101 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     5161 cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     5221 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     5281 taaactgttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     5341 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     5401 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     5461 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     5521 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggcg
     5581 agctctgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5641 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     5701 ctgcacctgc accagtcagt aaaacgacgg ccagtagtca aaagcctccg accggaggct
     5761 tttgacttgg ttcaggtgga gtgggag
//
