LOCUS       sequence-310685-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS186. 
            The crRNA-IS186 targets IS186 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-IS186
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     970..989
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1009..1086)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1132..1150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1151..1175
                     /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)."
     promoter        1419..1437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1438..1462
                     /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(6260..6278)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6274..6321
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6492..7283)
                     /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(7284..7375)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(8436..8510,1..1008))
                     /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"
ORIGIN
        1 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
       61 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
      121 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
      181 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
      241 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
      301 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
      361 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
      421 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
      481 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
      541 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
      601 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
      661 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
      721 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
      781 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
      841 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
      901 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
      961 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1021 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     1081 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac
     1141 tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt ttgtttaact
     1201 ttaataagga gatataccat gggtgaactg ccgagtaggt agctgataac tggatgcttt
     1261 gcgtgcaaag gaacctgaac tcgtgaactg ccgagtaggt agctgataac ggatccgaat
     1321 tcgagctcgg cgcgcctgca ggtcgacaag cttgcggccg cataatgctt aagtcgaaca
     1381 gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga
     1441 attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag aaggagatat
     1501 acatatgttt ttgcaaagac ctaaacctta cagcgatgaa agtttagaaa gtttctttat
     1561 ccgagtggct aacaaaaatg gctacggtga tgtccatcgc ttcctagaag ccactaaacg
     1621 attccttcaa gacattgacc ataatggcta tcaaaccttt ccgactgata taactcggat
     1681 aaacccatac tcagctaaaa acagttccag cgcacgaact gcgtcattcc tgaaacttgc
     1741 acaattgaca tttaatgaac cgccagagct acttgggttg gcaattaaca gaacaaacat
     1801 gaaatactcg ccgtcaacta gcgcggttgt tagaggtgca gaagtctttc ctcgcagttt
     1861 actacggacg cactccatcc cctgctgtcc tttgtgtctg cgagaaaatg gctacgcctc
     1921 ctacctttgg cactttcagg ggtacgaata ctgccacagc cataacgtac ctttaattac
     1981 cacttgtagc tgtggtaagg agtttgacta ccgagtatct gggttaaagg gcatttgctg
     2041 caaatgcaag gagcctatca ccttaaccag cagggagaac ggtcatgagg cagcgtgtac
     2101 tgtttcaaac tggcttgctg gccatgaatc taaacctctg ccaaatcttc ctaaaagcta
     2161 ccgatggggt ttagttcatt ggtggatggg tattaaagat agcgagttcg atcacttttc
     2221 gttcgttcaa tttttctcaa actggccaag gtcattccac tcgataatcg aagatgaagt
     2281 agagttcaac cttgagcatg ctgttgtcag cacgtctgaa ttacgactaa aagatcttct
     2341 tggtcgattg tttttcggtt caattcggtt acctgagcgg aatcttcaac acaatatcat
     2401 ccttggtgag cttctctgct atttagaaaa tcgtttatgg caagacaagg gattaatcgc
     2461 caacctcaaa atgaacgcgt tagaggcgac tgtaatgtta aattgtagcc tcgatcagat
     2521 tgcatcaatg gttgaacaac gcatcttgaa gccaaatcga aaaagcaagc ccaacagccc
     2581 tcttgatgtt accgattatc tatttcattt cggcgatatt ttctgtcttt ggttagctga
     2641 gttccaaagc gatgagttta accgttcgtt ttatgtgtcg aggtggtaaa tgcaaactct
     2701 gaaagaacta atcgcatcca atcctgacga cttaacaact gaacttaaga gagcatttcg
     2761 tccactcaca ccgcatattg caattgatgg taatgaactt gacgcactga cgatattagt
     2821 caatttaacc gataagactg atgatcagaa agacctgctc gatcgagcca aatgcaagca
     2881 aaaacttcga gatgaaaaat ggtgggctag ctgcataaat tgcgttaatt acagacaaag
     2941 ccataaccca aaattcccgg atatacgttc tgaaggcgtg attcgaaccc aagccctggg
     3001 tgaattaccg agtttcctac tctcgtcctc aaaaatccca ccataccatt ggtcatatag
     3061 ccatgattca aaatacgtca acaaaagcgc attcctcacc aatgagtttt gttgggatgg
     3121 tgagatctca tgtttgggtg agcttcttaa agatgcagat cacccacttt ggaatacttt
     3181 gaaaaagtta ggttgttctc aaaaaacttg caaagcaatg gcaaaacaac tagctgatat
     3241 tactctcacg actatcaatg tcacactcgc accaaattac ctgactcaaa tatctctccc
     3301 cgatagtgat acatcctaca tttcactctc tcctgttgca tcgctatcga tgcaaagcca
     3361 ctttcatcag aggctccaag atgaaaatcg gcatagcgcg ataacgcggt ttagccgaac
     3421 taccaacatg ggggtcactg cgatgacatg tggcggcgca tttagaatgt taaagtctgg
     3481 cgctaagttt tccagccccc ctcatcaccg attaaacagt aaacgaagtt ggttgacgtc
     3541 agagcatgtt cagtcattaa aacagtacca gcgcctcaac aaaagcctca tacctgaaaa
     3601 ctctcggatt gcactccgta gaaaatacaa aatcgagctt caaaatatgg tcagatcttg
     3661 gtttgcaatg caagaccata cccttgattc gaacatactt atccaacatt tgaatcatga
     3721 cctatcttac ttaggagcca caaaacgttt tgcatacgat ccagcgatga ccaagctctt
     3781 tactgagctt ttgaaacgag agttatcaaa ttcaatcaat aatggtgagc aacacactaa
     3841 tggatcgttt ttagtcctac cgaatatcag ggtttgtggc gcaacagctt taagctcccc
     3901 ggtaacggtg gggattccat cacttacagc tttctttggc ttcgttcacg catttgaacg
     3961 gaatataaat cgcaccacct catcgtttcg tgttgaatcc tttgcgatat gcgtccatca
     4021 actacatgtc gagaagcgag gtttgacagc agagtttgtg gaaaaaggcg acgggactat
     4081 atccgctccc gcgacccggg atgactggca gtgtgatgtc gtatttagcc ttattttgaa
     4141 caccaacttt gctcaacata ttgaccaaga tacgttagtt acatcactac caaagcgatt
     4201 ggctcggggt tcagcaaaaa ttgcgattga tgactttaaa catatcaact cattctcgac
     4261 attagaaaca gcgatcgaat ctctgccaat agaagctggt aggtggttat cactttacgc
     4321 acagtcaaac aataatctaa gtgatctatt agcagccatg acagaggacc atcagctcat
     4381 ggcaagctgc gtcggttacc acttgttaga agagcccaaa gataaaccaa actccctcag
     4441 aggttacaaa cacgctatcg ccgagtgcat cattggactc attaactcaa tcacctttag
     4501 ctcagagact gatcccaaca caatcttttg gtcgctaaag aactatcaaa actacctagt
     4561 ggtacagcca aggagtatca acgatgaaac taccgacaaa tctagcctat gagcgctcta
     4621 tcgacccatc agatgtctgt ttttttgtcg tctggcccga tgatagaaaa acacctttaa
     4681 cctacaattc tcgtactctg ctcgggcaaa tggaagcggc atcattagcc tatgatgtct
     4741 caggtcaacc aataaaaagt gccaccgctg aggcgttagc tcaagggaac cctcatcaag
     4801 ttgatttctg ccacgttcca tacggtgcga gtcatattga atgcagtttc tccgtctcgt
     4861 tttcttctga actacgtcaa ccatataagt gtaactcaag caaagttaaa caaacgctag
     4921 tgcaattagt cgagctctac gaaacgaaaa tcggctggac tgagctagca acccgatatt
     4981 tgatgaacat ttgcaacggt aaatggctgt ggaaaaatac ccgtaaagcc tattgctgga
     5041 acattgtact tacaccttgg ccttggaacg gggaaaaggt tggatttgaa gatattcgta
     5101 ctaactacac ctcacggcaa gactttaaaa ataataaaaa ttggtctgct atagttgaaa
     5161 tgatcaaaac cgcattttct agtactgatg ggctggcgat atttgaagtc agggccacct
     5221 tgcacttgcc aacgaatgct atggtgcggc caagccaagt tttcacagaa aaagaaagtg
     5281 gcagtaaaag taaatctaaa actcaaaaca gtcgagtttt tcagagtaca actattgatg
     5341 gtgaacgatc gccaatacta ggggccttta aaacgggagc agctattgca accattgacg
     5401 actggtatcc tgaagccact gagccactaa gggtcggacg gtttggggtt catcgcgaag
     5461 atgtcacttg ctaccgtcat ccgtctaccg gaaaagattt tttctcgata ttacaacaag
     5521 cagagcacta tattgaagtg ttgagcgcca acaaaactcc cgctcaagaa actatcaacg
     5581 acatgcactt tttaatggct aacctgatta agggtgggat gttccagcat aaaggagact
     5641 gactgtgaaa tggtattata agacaatcac ctttctgcca gagttgtgca acaacgagtc
     5701 actggctgca aagtgtctcc gcgttctgca tggatttaac tatcagtatg agacacgaaa
     5761 tatcggcgtt tcatttccgc tttggtgtga tgcaacggtt ggaaaaaaga tttcatttgt
     5821 cagcaagaac aagatagaac tcgacttact acttaaacaa cactatttcg tccaaatgga
     5881 acaacttcaa tactttcata tatccaacac tgttctcgtc ccagaagatt gtacatacgt
     5941 ttcctttaga cgctgtcaat ctatagataa gctcacagca gcagggctgg caaggaaaat
     6001 cagacgcctg gagaaacgtg ctctatctag aggcgagcaa tttgacccat catcttttgc
     6061 tcaaaaagag catactgcaa tagcgcacta ccactcactt ggggagtcca gcaaacagac
     6121 gaaccgcaac tttcgactca atatcaggat gctctcggag caaccccgtg aggggaactc
     6181 gatttttagt agctatggct tatcaaattc agaaaactcg tttcagcctg tacccttaat
     6241 ctgacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     6301 cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg
     6361 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
     6421 tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga
     6481 attgttagac attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     6541 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     6601 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     6661 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     6721 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     6781 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     6841 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     6901 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     6961 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     7021 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     7081 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     7141 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     7201 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     7261 tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag catttatcag
     7321 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaatagct
     7381 agctcactcg gtcgctacgc tccgggcgtg agactgcggc gggcgctgcg gacacataca
     7441 aagttaccca cagattccgt ggataagcag gggactaaca tgtgaggcaa aacagcaggg
     7501 ccgcgccggt ggcgtttttc cataggctcc gccctcctgc cagagttcac ataaacagac
     7561 gcttttccgg tgcatctgtg ggagccgtga ggctcaacca tgaatctgac agtacgggcg
     7621 aaacccgaca ggacttaaag atccccaccg tttccggcgg gtcgctccct cttgcgctct
     7681 cctgttccga ccctgccgtt taccggatac ctgttccgcc tttctccctt acgggaagtg
     7741 tggcgctttc tcatagctca cacactggta tctcggctcg gtgtaggtcg ttcgctccaa
     7801 gctgggctgt aagcaagaac tccccgttca gcccgactgc tgcgccttat ccggtaactg
     7861 ttcacttgag tccaacccgg aaaagcacgg taaaacgcca ctggcagcag ccattggtaa
     7921 ctgggagttc gcagaggatt tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca
     7981 aagtccggct acactggaag gacagatttg gttgctgtgc tctgcgaaag ccagttacca
     8041 cggttaagca gttccccaac tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt
     8101 ttcgaaggtt aagtcagttg gggaactgct taaccttcga tcaaaccacc tccccaggtg
     8161 gttttttcgt ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct
     8221 ttgatctttt ctactgaacc gctctagatt tcagtgcaat ttatctcttc aaatgtagca
     8281 cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc
     8341 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     8401 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
//
