LOCUS       sequence-305981-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-tra13. 
            The crRNA-tra13 targets 13 copies transposon loci in Tatumella 
            citrea..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8510)
  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 8510)
  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: pQCascade-tra13
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        73..91
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    92..116
                     /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     complement(4914..4932)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4928..4975
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5146..5937)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5938..6029)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7090..8172)
                     /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     8134..8153
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8173..8250)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        8296..8314
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8315..8339
                     /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)."
ORIGIN
        1 caagcttgcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg tacacggccg
       61 cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccat
      121 cttagtatat tagttaagta taagaaggag atatacatat gtttttgcaa agacctaaac
      181 cttacagcga tgaaagttta gaaagtttct ttatccgagt ggctaacaaa aatggctacg
      241 gtgatgtcca tcgcttccta gaagccacta aacgattcct tcaagacatt gaccataatg
      301 gctatcaaac ctttccgact gatataactc ggataaaccc atactcagct aaaaacagtt
      361 ccagcgcacg aactgcgtca ttcctgaaac ttgcacaatt gacatttaat gaaccgccag
      421 agctacttgg gttggcaatt aacagaacaa acatgaaata ctcgccgtca actagcgcgg
      481 ttgttagagg tgcagaagtc tttcctcgca gtttactacg gacgcactcc atcccctgct
      541 gtcctttgtg tctgcgagaa aatggctacg cctcctacct ttggcacttt caggggtacg
      601 aatactgcca cagccataac gtacctttaa ttaccacttg tagctgtggt aaggagtttg
      661 actaccgagt atctgggtta aagggcattt gctgcaaatg caaggagcct atcaccttaa
      721 ccagcaggga gaacggtcat gaggcagcgt gtactgtttc aaactggctt gctggccatg
      781 aatctaaacc tctgccaaat cttcctaaaa gctaccgatg gggtttagtt cattggtgga
      841 tgggtattaa agatagcgag ttcgatcact tttcgttcgt tcaatttttc tcaaactggc
      901 caaggtcatt ccactcgata atcgaagatg aagtagagtt caaccttgag catgctgttg
      961 tcagcacgtc tgaattacga ctaaaagatc ttcttggtcg attgtttttc ggttcaattc
     1021 ggttacctga gcggaatctt caacacaata tcatccttgg tgagcttctc tgctatttag
     1081 aaaatcgttt atggcaagac aagggattaa tcgccaacct caaaatgaac gcgttagagg
     1141 cgactgtaat gttaaattgt agcctcgatc agattgcatc aatggttgaa caacgcatct
     1201 tgaagccaaa tcgaaaaagc aagcccaaca gccctcttga tgttaccgat tatctatttc
     1261 atttcggcga tattttctgt ctttggttag ctgagttcca aagcgatgag tttaaccgtt
     1321 cgttttatgt gtcgaggtgg taaatgcaaa ctctgaaaga actaatcgca tccaatcctg
     1381 acgacttaac aactgaactt aagagagcat ttcgtccact cacaccgcat attgcaattg
     1441 atggtaatga acttgacgca ctgacgatat tagtcaattt aaccgataag actgatgatc
     1501 agaaagacct gctcgatcga gccaaatgca agcaaaaact tcgagatgaa aaatggtggg
     1561 ctagctgcat aaattgcgtt aattacagac aaagccataa cccaaaattc ccggatatac
     1621 gttctgaagg cgtgattcga acccaagccc tgggtgaatt accgagtttc ctactctcgt
     1681 cctcaaaaat cccaccatac cattggtcat atagccatga ttcaaaatac gtcaacaaaa
     1741 gcgcattcct caccaatgag ttttgttggg atggtgagat ctcatgtttg ggtgagcttc
     1801 ttaaagatgc agatcaccca ctttggaata ctttgaaaaa gttaggttgt tctcaaaaaa
     1861 cttgcaaagc aatggcaaaa caactagctg atattactct cacgactatc aatgtcacac
     1921 tcgcaccaaa ttacctgact caaatatctc tccccgatag tgatacatcc tacatttcac
     1981 tctctcctgt tgcatcgcta tcgatgcaaa gccactttca tcagaggctc caagatgaaa
     2041 atcggcatag cgcgataacg cggtttagcc gaactaccaa catgggggtc actgcgatga
     2101 catgtggcgg cgcatttaga atgttaaagt ctggcgctaa gttttccagc ccccctcatc
     2161 accgattaaa cagtaaacga agttggttga cgtcagagca tgttcagtca ttaaaacagt
     2221 accagcgcct caacaaaagc ctcatacctg aaaactctcg gattgcactc cgtagaaaat
     2281 acaaaatcga gcttcaaaat atggtcagat cttggtttgc aatgcaagac catacccttg
     2341 attcgaacat acttatccaa catttgaatc atgacctatc ttacttagga gccacaaaac
     2401 gttttgcata cgatccagcg atgaccaagc tctttactga gcttttgaaa cgagagttat
     2461 caaattcaat caataatggt gagcaacaca ctaatggatc gtttttagtc ctaccgaata
     2521 tcagggtttg tggcgcaaca gctttaagct ccccggtaac ggtggggatt ccatcactta
     2581 cagctttctt tggcttcgtt cacgcatttg aacggaatat aaatcgcacc acctcatcgt
     2641 ttcgtgttga atcctttgcg atatgcgtcc atcaactaca tgtcgagaag cgaggtttga
     2701 cagcagagtt tgtggaaaaa ggcgacggga ctatatccgc tcccgcgacc cgggatgact
     2761 ggcagtgtga tgtcgtattt agccttattt tgaacaccaa ctttgctcaa catattgacc
     2821 aagatacgtt agttacatca ctaccaaagc gattggctcg gggttcagca aaaattgcga
     2881 ttgatgactt taaacatatc aactcattct cgacattaga aacagcgatc gaatctctgc
     2941 caatagaagc tggtaggtgg ttatcacttt acgcacagtc aaacaataat ctaagtgatc
     3001 tattagcagc catgacagag gaccatcagc tcatggcaag ctgcgtcggt taccacttgt
     3061 tagaagagcc caaagataaa ccaaactccc tcagaggtta caaacacgct atcgccgagt
     3121 gcatcattgg actcattaac tcaatcacct ttagctcaga gactgatccc aacacaatct
     3181 tttggtcgct aaagaactat caaaactacc tagtggtaca gccaaggagt atcaacgatg
     3241 aaactaccga caaatctagc ctatgagcgc tctatcgacc catcagatgt ctgttttttt
     3301 gtcgtctggc ccgatgatag aaaaacacct ttaacctaca attctcgtac tctgctcggg
     3361 caaatggaag cggcatcatt agcctatgat gtctcaggtc aaccaataaa aagtgccacc
     3421 gctgaggcgt tagctcaagg gaaccctcat caagttgatt tctgccacgt tccatacggt
     3481 gcgagtcata ttgaatgcag tttctccgtc tcgttttctt ctgaactacg tcaaccatat
     3541 aagtgtaact caagcaaagt taaacaaacg ctagtgcaat tagtcgagct ctacgaaacg
     3601 aaaatcggct ggactgagct agcaacccga tatttgatga acatttgcaa cggtaaatgg
     3661 ctgtggaaaa atacccgtaa agcctattgc tggaacattg tacttacacc ttggccttgg
     3721 aacggggaaa aggttggatt tgaagatatt cgtactaact acacctcacg gcaagacttt
     3781 aaaaataata aaaattggtc tgctatagtt gaaatgatca aaaccgcatt ttctagtact
     3841 gatgggctgg cgatatttga agtcagggcc accttgcact tgccaacgaa tgctatggtg
     3901 cggccaagcc aagttttcac agaaaaagaa agtggcagta aaagtaaatc taaaactcaa
     3961 aacagtcgag tttttcagag tacaactatt gatggtgaac gatcgccaat actaggggcc
     4021 tttaaaacgg gagcagctat tgcaaccatt gacgactggt atcctgaagc cactgagcca
     4081 ctaagggtcg gacggtttgg ggttcatcgc gaagatgtca cttgctaccg tcatccgtct
     4141 accggaaaag attttttctc gatattacaa caagcagagc actatattga agtgttgagc
     4201 gccaacaaaa ctcccgctca agaaactatc aacgacatgc actttttaat ggctaacctg
     4261 attaagggtg ggatgttcca gcataaagga gactgactgt gaaatggtat tataagacaa
     4321 tcacctttct gccagagttg tgcaacaacg agtcactggc tgcaaagtgt ctccgcgttc
     4381 tgcatggatt taactatcag tatgagacac gaaatatcgg cgtttcattt ccgctttggt
     4441 gtgatgcaac ggttggaaaa aagatttcat ttgtcagcaa gaacaagata gaactcgact
     4501 tactacttaa acaacactat ttcgtccaaa tggaacaact tcaatacttt catatatcca
     4561 acactgttct cgtcccagaa gattgtacat acgtttcctt tagacgctgt caatctatag
     4621 ataagctcac agcagcaggg ctggcaagga aaatcagacg cctggagaaa cgtgctctat
     4681 ctagaggcga gcaatttgac ccatcatctt ttgctcaaaa agagcatact gcaatagcgc
     4741 actaccactc acttggggag tccagcaaac agacgaaccg caactttcga ctcaatatca
     4801 ggatgctctc ggagcaaccc cgtgagggga actcgatttt tagtagctat ggcttatcaa
     4861 attcagaaaa ctcgtttcag cctgtaccct taatctgacc taggctgctg ccaccgctga
     4921 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     4981 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     5041 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcatc
     5101 gtggccggat cttgcggccc ctcggcttga acgaattgtt agacattatt tgccgactac
     5161 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     5221 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     5281 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     5341 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     5401 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     5461 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     5521 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     5581 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     5641 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     5701 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     5761 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     5821 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     5881 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatact
     5941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6001 atttgaatgt atttagaaaa ataaacaaat agctagctca ctcggtcgct acgctccggg
     6061 cgtgagactg cggcgggcgc tgcggacaca tacaaagtta cccacagatt ccgtggataa
     6121 gcaggggact aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt tttccatagg
     6181 ctccgccctc ctgccagagt tcacataaac agacgctttt ccggtgcatc tgtgggagcc
     6241 gtgaggctca accatgaatc tgacagtacg ggcgaaaccc gacaggactt aaagatcccc
     6301 accgtttccg gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg
     6361 atacctgttc cgcctttctc ccttacggga agtgtggcgc tttctcatag ctcacacact
     6421 ggtatctcgg ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg
     6481 ttcagcccga ctgctgcgcc ttatccggta actgttcact tgagtccaac ccggaaaagc
     6541 acggtaaaac gccactggca gcagccattg gtaactggga gttcgcagag gatttgttta
     6601 gctaaacacg cggttgctct tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga
     6661 tttggttgct gtgctctgcg aaagccagtt accacggtta agcagttccc caactgactt
     6721 aaccttcgat caaaccacct ccccaggtgg ttttttcgaa ggttaagtca gttggggaac
     6781 tgcttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat
     6841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta
     6901 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
     6961 gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     7021 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     7081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7141 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     7201 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     7261 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     7321 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
     7381 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     7441 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     7501 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     7561 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     7621 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     7681 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     7741 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     7801 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     7861 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     7921 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     7981 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     8041 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     8101 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     8161 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     8221 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     8281 cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     8341 ctgtagaaat aattttgttt aactttaata aggagatata ccatgggtga actgccgagt
     8401 aggtagctga taactgaaaa tgcagaggcc ttcggcaaag catctagtga actgccgagt
     8461 aggtagctga taacggatcc gaattcgagc tcggcgcgcc tgcaggtcga
//
