LOCUS       sequence-309888-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TnsABC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  AUTHORS   Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian 
            Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
  TITLE     Multicopy chromosomal integration using CRISPR-associated 
            transposases.
  JOURNAL   ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
  PUBMED    32551502
REFERENCE   2  (bases 1 to 6858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073."
COMMENT     Sequence Label: pTnsABC
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             2149..2160
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3587..3605)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3601..3648
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3878..4693)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3878..4693)
                     /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"
     primer_bind     4601..4620
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4694..4785)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4803..5438)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(4948..4967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5636..6718)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6680..6699
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6719..6796)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6842..6858,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6842..6858,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcgacaa gtttacctac gccctcagca attacgactt cggcgttaga
      121 gtatgcattc catactcccg ctcgcaatct aacgaaatct cgcggaaaaa acattcatcg
      181 ttatgtcagt gtaaagatga gtaagaggat tacggtagaa tctactctag agtgtgatgc
      241 ctgctatcac tttgattttg agccaagtat tgttcgcttt tgcgctcaac cgattcgatt
      301 tttatattat ctcaatggtc agtctcactc ctatgttcct gactttctag ttcaatttga
      361 taccaacgag tttgttctat atgaagtaaa gtcagcttat gctaagaaca aacctgattt
      421 tgatgttgaa tgggaggcga aagtaaaagc agcaactgaa ctagggctag aattggagct
      481 tgttgaagag agtgatatta gggatacggt tgtattaaat aatcttaagc gcatgcatcg
      541 ttatgcttcg aaagatgagc tgaataacgt acataactct ctcttaaaaa taataaagta
      601 caatggcgcc caatctgcaa gatgcttggg agaacagttg ggtttaaaag gccgaactgt
      661 tttaccaatt ttgtgcgatt tgctgtcaag gtgtttactc gatacacgtt tggataagcc
      721 tctatctctt gaatctcgat ttgagttggc cagttatggc taagaaaggg ttctcaagtt
      781 tccatagaaa agcagtctct tcccaagata cgcttgaatc gatagagctt gtctctagcg
      841 ctaattgtct agaaagtgtt acgtatcaag atatatcagc atttcccgaa acaattgcgg
      901 tagagattaa tttccgatta agcattcttc gctttttagc gcgaaagtgc gaaaccattg
      961 tggccaaatc aattgaacca catcgtgtag agctacagca aaactatagt agaaaaatac
     1021 ccagtgcaat aacgatatat cgatggtggc ttgcttttcg aaaatcagac tacaacccca
     1081 ttagcttagc acctaatatc aaggatagag gtaatagaga gacaaaagtg tcaacagttg
     1141 ttgattctat tatggaacag gcagttgaaa gagttatatc tggacgaaaa gtcaatgtta
     1201 gctctgcata taaacgtgtt cgacgaaaag ttcgtcaata caatctaact catggaacga
     1261 aatacacgta tcctaagtac gaatctgtaa gaaagcgagt aaaaaagaaa accccatttg
     1321 agttattagc cgcagggaaa ggggagagag tagctaagag agagtttcgc cgaatgggaa
     1381 aaaagatcct cacgtctagc gtgctagaga gggttgaaat agatcacact gtcgttgacc
     1441 tttttgcagt acacgaagag tatcgaatcc cattgggccg accttggctt actcaattgg
     1501 ttgattgtta cagtaaagct gtaatcggtt tttatttagg tttcgagcct cctagctatg
     1561 tgtcggtttc ccttgcactt aagaatgcaa tacaacgcaa agatgactta atctcctcgt
     1621 atgaatcgat cgagaatgaa tggctatgtt atggcatccc agacctactc gtaactgata
     1681 atggtaaaga gtttttgtcg aaagcatttg atcaagcatg tgaatcacta ttgatcaatg
     1741 tgcatcaaaa taaagttgag acgcccgaca acaaacctca tgttgaacgt aactacggga
     1801 ctattaatac ttctctgtta gacgatttac ctgggaaatc cttcagccag taccttcaaa
     1861 gagaagggta cgactctgtg ggagaagcta cccttacact caatgagatt agagaaattt
     1921 acttaatttg gttggtggat atttatcata aaaaacccaa tcagagaggc actaattgtc
     1981 ctaatgttgc gtggaaaaag ggttgtcaag aatgggaacc agaggagttc tctggttcta
     2041 aagacgaatt agactttaaa tttgctattg ttgattacaa acaacttact aaagtaggga
     2101 taactgtcta caaagaactg agttatagca atgaccgttt agctgaatat agagggaaga
     2161 aaggaaacca taaagttcag ttcaagtata accctgagtg tatggcagtt atttgggtgt
     2221 tggatgagga tatgaatgag tactttacag ttaatgcgat tgactacgaa tatgcaagta
     2281 gagtatcact ttggcaacat aaatataaca tgaaatatca agcagaacta aattcagcag
     2341 aatatgatga ggacaaggaa attgatgcag aaataaaaat tgaagaaatc gcagatcgtt
     2401 caattgttaa gactaacaaa atcagagctc ggaggcgtgg cgctaggcat caagagaata
     2461 gcgcaagggc taagtcaatc agtaatgcga acccggcctc gatacaaaaa catgaagatg
     2521 aaatcgttag tgcagataat gacgattggg atattgatta tgtctgagaa atcgtcaaat
     2581 gagtgaaacg cgtgaggctc gtatatcaag agctaaaagg gcatttgtat ccacaccgag
     2641 cgttaggaaa atcttgagtt acatggatag atgtagagat ctatcagacc tagagtctga
     2701 gcctacatgc atgatggtct atggtgcttc gggtgtaggt aaaacgaccg tcatcaagaa
     2761 atacttaaat caaaacagaa gagagtccga agccgggggc gatataatac cggttttgca
     2821 tattgagttg ccagacaatg cgaagccagt agatgcagca agggaattgc tggttgaaat
     2881 gggtgacccg ctagcacttt atgaaactga cttagctaga ttgacgaaaa gactgactga
     2941 attaatccct gcggtcggcg tgaagctgat tattatcgat gagttccaac atttggtgga
     3001 agaaaggtca aatcgggttc ttacccaagt aggtaattgg ctaaaaatga tacttaacaa
     3061 aacgaaatgt ccaattgtta tatttggtat gccatactca aaagttgtac tgcaagcaaa
     3121 ctcgcaactt cacgggcgat tttccattca ggttgaactg cgccccttta gctaccaggg
     3181 aggtagaggt gtatttaaaa cttttttgga ataccttgat aaagccctac cttttgaaaa
     3241 acaggctggc ttagccaacg aaagtttgca gaaaaaattg tatgcattct ctcagggaaa
     3301 catgcgttcg ttgagaaacc ttatttatca agcatctatc gaagcaattg ataatcagca
     3361 tgagacgata accgaagaag atttcgtttt tgcatcgaag ttgacatcgg gcgataaacc
     3421 caactcatgg aaaaatcctt ttgaggaggg tgttgaggta acagaagata tgttacgacc
     3481 gccaccaaaa gatattggtt gggaggacta tttgagacat tcaaccccga gagtgagtaa
     3541 accaggtaga aataaaaact ttttcgaata acctaggctg ctgccaccgc tgagcaataa
     3601 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     3661 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     3721 tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt
     3781 ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3841 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     3901 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3961 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     4021 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     4081 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     4141 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     4201 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     4261 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     4321 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     4381 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     4441 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     4501 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     4561 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     4621 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     4681 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc atgctagcgc
     4801 agaaacgtcc tagaagatgc caggaggata cttagcagag agacaataag gccggagcga
     4861 agccgttttt ccataggctc cgcccccctg acgaacatca cgaaatctga cgctcaaatc
     4921 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct gatggctccc
     4981 tcttgcgctc tcctgttccc gtcctgcggc gtccgtgttg tggtggaggc tttacccaaa
     5041 tcaccacgtc ccgttccgtg tagacagttc gctccaagct gggctgtgtg caagaacccc
     5101 ccgttcagcc cgactgctgc gccttatccg gtaactatca tcttgagtcc aacccggaaa
     5161 gacacgacaa aacgccactg gcagcagcca ttggtaactg agaattagtg gatttagata
     5221 tcgagagtct tgaagtggtg gcctaacaga ggctacactg aaaggacagt atttggtatc
     5281 tgcgctccac taaagccagt taccaggtta agcagttccc caactgactt aaccttcgat
     5341 caaaccgcct ccccaggcgg ttttttcgtt tacagagcag gagattacga cgatcgtaaa
     5401 aggatctcaa gaagatcctt tacggattcc cgacaccatc actctagatt tcagtgcaat
     5461 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     5521 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5581 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     5641 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5701 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     5761 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     5821 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     5881 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
     5941 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     6001 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     6061 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     6121 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     6181 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     6241 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     6301 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     6361 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     6421 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     6481 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     6541 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     6601 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     6661 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     6721 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     6781 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa
     6841 ttaatacgac tcactata
//
