LOCUS       sequence-305996-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS2. The 
            crRNA-IS2 targets IS2 loci in Escherchia coli..
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-IS2
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        166..184
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    185..209
                     /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(5007..5025)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5021..5068
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5239..6030)
                     /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(6031..6122)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7183..8265)
                     /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     8227..8246
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8266..8343)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        8389..8407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8408..8432
                     /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 cagaggcgtt cgagcattat aacgaatggg tgaactgccg agtaggtagc tgataacgga
       61 tccgaattcg agctcggcgc gcctgcaggt cgacaagctt gcggccgcat aatgcttaag
      121 tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata cgactcacta
      181 taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa gtataagaag
      241 gagatataca tatgtttttg caaagaccta aaccttacag cgatgaaagt ttagaaagtt
      301 tctttatccg agtggctaac aaaaatggct acggtgatgt ccatcgcttc ctagaagcca
      361 ctaaacgatt ccttcaagac attgaccata atggctatca aacctttccg actgatataa
      421 ctcggataaa cccatactca gctaaaaaca gttccagcgc acgaactgcg tcattcctga
      481 aacttgcaca attgacattt aatgaaccgc cagagctact tgggttggca attaacagaa
      541 caaacatgaa atactcgccg tcaactagcg cggttgttag aggtgcagaa gtctttcctc
      601 gcagtttact acggacgcac tccatcccct gctgtccttt gtgtctgcga gaaaatggct
      661 acgcctccta cctttggcac tttcaggggt acgaatactg ccacagccat aacgtacctt
      721 taattaccac ttgtagctgt ggtaaggagt ttgactaccg agtatctggg ttaaagggca
      781 tttgctgcaa atgcaaggag cctatcacct taaccagcag ggagaacggt catgaggcag
      841 cgtgtactgt ttcaaactgg cttgctggcc atgaatctaa acctctgcca aatcttccta
      901 aaagctaccg atggggttta gttcattggt ggatgggtat taaagatagc gagttcgatc
      961 acttttcgtt cgttcaattt ttctcaaact ggccaaggtc attccactcg ataatcgaag
     1021 atgaagtaga gttcaacctt gagcatgctg ttgtcagcac gtctgaatta cgactaaaag
     1081 atcttcttgg tcgattgttt ttcggttcaa ttcggttacc tgagcggaat cttcaacaca
     1141 atatcatcct tggtgagctt ctctgctatt tagaaaatcg tttatggcaa gacaagggat
     1201 taatcgccaa cctcaaaatg aacgcgttag aggcgactgt aatgttaaat tgtagcctcg
     1261 atcagattgc atcaatggtt gaacaacgca tcttgaagcc aaatcgaaaa agcaagccca
     1321 acagccctct tgatgttacc gattatctat ttcatttcgg cgatattttc tgtctttggt
     1381 tagctgagtt ccaaagcgat gagtttaacc gttcgtttta tgtgtcgagg tggtaaatgc
     1441 aaactctgaa agaactaatc gcatccaatc ctgacgactt aacaactgaa cttaagagag
     1501 catttcgtcc actcacaccg catattgcaa ttgatggtaa tgaacttgac gcactgacga
     1561 tattagtcaa tttaaccgat aagactgatg atcagaaaga cctgctcgat cgagccaaat
     1621 gcaagcaaaa acttcgagat gaaaaatggt gggctagctg cataaattgc gttaattaca
     1681 gacaaagcca taacccaaaa ttcccggata tacgttctga aggcgtgatt cgaacccaag
     1741 ccctgggtga attaccgagt ttcctactct cgtcctcaaa aatcccacca taccattggt
     1801 catatagcca tgattcaaaa tacgtcaaca aaagcgcatt cctcaccaat gagttttgtt
     1861 gggatggtga gatctcatgt ttgggtgagc ttcttaaaga tgcagatcac ccactttgga
     1921 atactttgaa aaagttaggt tgttctcaaa aaacttgcaa agcaatggca aaacaactag
     1981 ctgatattac tctcacgact atcaatgtca cactcgcacc aaattacctg actcaaatat
     2041 ctctccccga tagtgataca tcctacattt cactctctcc tgttgcatcg ctatcgatgc
     2101 aaagccactt tcatcagagg ctccaagatg aaaatcggca tagcgcgata acgcggttta
     2161 gccgaactac caacatgggg gtcactgcga tgacatgtgg cggcgcattt agaatgttaa
     2221 agtctggcgc taagttttcc agcccccctc atcaccgatt aaacagtaaa cgaagttggt
     2281 tgacgtcaga gcatgttcag tcattaaaac agtaccagcg cctcaacaaa agcctcatac
     2341 ctgaaaactc tcggattgca ctccgtagaa aatacaaaat cgagcttcaa aatatggtca
     2401 gatcttggtt tgcaatgcaa gaccataccc ttgattcgaa catacttatc caacatttga
     2461 atcatgacct atcttactta ggagccacaa aacgttttgc atacgatcca gcgatgacca
     2521 agctctttac tgagcttttg aaacgagagt tatcaaattc aatcaataat ggtgagcaac
     2581 acactaatgg atcgttttta gtcctaccga atatcagggt ttgtggcgca acagctttaa
     2641 gctccccggt aacggtgggg attccatcac ttacagcttt ctttggcttc gttcacgcat
     2701 ttgaacggaa tataaatcgc accacctcat cgtttcgtgt tgaatccttt gcgatatgcg
     2761 tccatcaact acatgtcgag aagcgaggtt tgacagcaga gtttgtggaa aaaggcgacg
     2821 ggactatatc cgctcccgcg acccgggatg actggcagtg tgatgtcgta tttagcctta
     2881 ttttgaacac caactttgct caacatattg accaagatac gttagttaca tcactaccaa
     2941 agcgattggc tcggggttca gcaaaaattg cgattgatga ctttaaacat atcaactcat
     3001 tctcgacatt agaaacagcg atcgaatctc tgccaataga agctggtagg tggttatcac
     3061 tttacgcaca gtcaaacaat aatctaagtg atctattagc agccatgaca gaggaccatc
     3121 agctcatggc aagctgcgtc ggttaccact tgttagaaga gcccaaagat aaaccaaact
     3181 ccctcagagg ttacaaacac gctatcgccg agtgcatcat tggactcatt aactcaatca
     3241 cctttagctc agagactgat cccaacacaa tcttttggtc gctaaagaac tatcaaaact
     3301 acctagtggt acagccaagg agtatcaacg atgaaactac cgacaaatct agcctatgag
     3361 cgctctatcg acccatcaga tgtctgtttt tttgtcgtct ggcccgatga tagaaaaaca
     3421 cctttaacct acaattctcg tactctgctc gggcaaatgg aagcggcatc attagcctat
     3481 gatgtctcag gtcaaccaat aaaaagtgcc accgctgagg cgttagctca agggaaccct
     3541 catcaagttg atttctgcca cgttccatac ggtgcgagtc atattgaatg cagtttctcc
     3601 gtctcgtttt cttctgaact acgtcaacca tataagtgta actcaagcaa agttaaacaa
     3661 acgctagtgc aattagtcga gctctacgaa acgaaaatcg gctggactga gctagcaacc
     3721 cgatatttga tgaacatttg caacggtaaa tggctgtgga aaaatacccg taaagcctat
     3781 tgctggaaca ttgtacttac accttggcct tggaacgggg aaaaggttgg atttgaagat
     3841 attcgtacta actacacctc acggcaagac tttaaaaata ataaaaattg gtctgctata
     3901 gttgaaatga tcaaaaccgc attttctagt actgatgggc tggcgatatt tgaagtcagg
     3961 gccaccttgc acttgccaac gaatgctatg gtgcggccaa gccaagtttt cacagaaaaa
     4021 gaaagtggca gtaaaagtaa atctaaaact caaaacagtc gagtttttca gagtacaact
     4081 attgatggtg aacgatcgcc aatactaggg gcctttaaaa cgggagcagc tattgcaacc
     4141 attgacgact ggtatcctga agccactgag ccactaaggg tcggacggtt tggggttcat
     4201 cgcgaagatg tcacttgcta ccgtcatccg tctaccggaa aagatttttt ctcgatatta
     4261 caacaagcag agcactatat tgaagtgttg agcgccaaca aaactcccgc tcaagaaact
     4321 atcaacgaca tgcacttttt aatggctaac ctgattaagg gtgggatgtt ccagcataaa
     4381 ggagactgac tgtgaaatgg tattataaga caatcacctt tctgccagag ttgtgcaaca
     4441 acgagtcact ggctgcaaag tgtctccgcg ttctgcatgg atttaactat cagtatgaga
     4501 cacgaaatat cggcgtttca tttccgcttt ggtgtgatgc aacggttgga aaaaagattt
     4561 catttgtcag caagaacaag atagaactcg acttactact taaacaacac tatttcgtcc
     4621 aaatggaaca acttcaatac tttcatatat ccaacactgt tctcgtccca gaagattgta
     4681 catacgtttc ctttagacgc tgtcaatcta tagataagct cacagcagca gggctggcaa
     4741 ggaaaatcag acgcctggag aaacgtgctc tatctagagg cgagcaattt gacccatcat
     4801 cttttgctca aaaagagcat actgcaatag cgcactacca ctcacttggg gagtccagca
     4861 aacagacgaa ccgcaacttt cgactcaata tcaggatgct ctcggagcaa ccccgtgagg
     4921 ggaactcgat ttttagtagc tatggcttat caaattcaga aaactcgttt cagcctgtac
     4981 ccttaatctg acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     5041 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
     5101 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
     5161 acgaccctgc cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct
     5221 tgaacgaatt gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     5281 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
     5341 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     5401 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     5461 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
     5521 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     5581 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     5641 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     5701 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     5761 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     5821 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     5881 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     5941 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     6001 cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat
     6061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6121 aatagctagc tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac
     6181 acatacaaag ttacccacag attccgtgga taagcagggg actaacatgt gaggcaaaac
     6241 agcagggccg cgccggtggc gtttttccat aggctccgcc ctcctgccag agttcacata
     6301 aacagacgct tttccggtgc atctgtggga gccgtgaggc tcaaccatga atctgacagt
     6361 acgggcgaaa cccgacagga cttaaagatc cccaccgttt ccggcgggtc gctccctctt
     6421 gcgctctcct gttccgaccc tgccgtttac cggatacctg ttccgccttt ctcccttacg
     6481 ggaagtgtgg cgctttctca tagctcacac actggtatct cggctcggtg taggtcgttc
     6541 gctccaagct gggctgtaag caagaactcc ccgttcagcc cgactgctgc gccttatccg
     6601 gtaactgttc acttgagtcc aacccggaaa agcacggtaa aacgccactg gcagcagcca
     6661 ttggtaactg ggagttcgca gaggatttgt ttagctaaac acgcggttgc tcttgaagtg
     6721 tgcgccaaag tccggctaca ctggaaggac agatttggtt gctgtgctct gcgaaagcca
     6781 gttaccacgg ttaagcagtt ccccaactga cttaaccttc gatcaaacca cctccccagg
     6841 tggttttttc gaaggttaag tcagttgggg aactgcttaa ccttcgatca aaccacctcc
     6901 ccaggtggtt ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga
     6961 agatcctttg atcttttcta ctgaaccgct ctagatttca gtgcaattta tctcttcaaa
     7021 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
     7081 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     7141 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     7201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7261 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     7321 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     7381 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     7441 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     7501 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     7561 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     7621 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     7681 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     7741 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     7801 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     7861 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     7921 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     7981 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     8041 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     8101 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     8161 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     8221 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     8281 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     8341 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
     8401 ctatagggga attgtgagcg gataacaatt cccctgtaga aataattttg tttaacttta
     8461 ataaggagat ataccatggg tgaactgccg agtaggtagc tgataacttg
//
