LOCUS       sequence-305994-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS3. The 
            crRNA-IS3 targets IS3 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-IS3
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        119..137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    138..162
                     /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(4960..4978)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4974..5021
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5192..5983)
                     /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(5984..6075)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7136..8218)
                     /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     8180..8199
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8219..8296)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        8342..8360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8361..8385
                     /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 agctgataac ggatccgaat tcgagctcgg cgcgcctgca ggtcgacaag cttgcggccg
       61 cataatgctt aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta
      121 atacgactca ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt
      181 taagtataag aaggagatat acatatgttt ttgcaaagac ctaaacctta cagcgatgaa
      241 agtttagaaa gtttctttat ccgagtggct aacaaaaatg gctacggtga tgtccatcgc
      301 ttcctagaag ccactaaacg attccttcaa gacattgacc ataatggcta tcaaaccttt
      361 ccgactgata taactcggat aaacccatac tcagctaaaa acagttccag cgcacgaact
      421 gcgtcattcc tgaaacttgc acaattgaca tttaatgaac cgccagagct acttgggttg
      481 gcaattaaca gaacaaacat gaaatactcg ccgtcaacta gcgcggttgt tagaggtgca
      541 gaagtctttc ctcgcagttt actacggacg cactccatcc cctgctgtcc tttgtgtctg
      601 cgagaaaatg gctacgcctc ctacctttgg cactttcagg ggtacgaata ctgccacagc
      661 cataacgtac ctttaattac cacttgtagc tgtggtaagg agtttgacta ccgagtatct
      721 gggttaaagg gcatttgctg caaatgcaag gagcctatca ccttaaccag cagggagaac
      781 ggtcatgagg cagcgtgtac tgtttcaaac tggcttgctg gccatgaatc taaacctctg
      841 ccaaatcttc ctaaaagcta ccgatggggt ttagttcatt ggtggatggg tattaaagat
      901 agcgagttcg atcacttttc gttcgttcaa tttttctcaa actggccaag gtcattccac
      961 tcgataatcg aagatgaagt agagttcaac cttgagcatg ctgttgtcag cacgtctgaa
     1021 ttacgactaa aagatcttct tggtcgattg tttttcggtt caattcggtt acctgagcgg
     1081 aatcttcaac acaatatcat ccttggtgag cttctctgct atttagaaaa tcgtttatgg
     1141 caagacaagg gattaatcgc caacctcaaa atgaacgcgt tagaggcgac tgtaatgtta
     1201 aattgtagcc tcgatcagat tgcatcaatg gttgaacaac gcatcttgaa gccaaatcga
     1261 aaaagcaagc ccaacagccc tcttgatgtt accgattatc tatttcattt cggcgatatt
     1321 ttctgtcttt ggttagctga gttccaaagc gatgagttta accgttcgtt ttatgtgtcg
     1381 aggtggtaaa tgcaaactct gaaagaacta atcgcatcca atcctgacga cttaacaact
     1441 gaacttaaga gagcatttcg tccactcaca ccgcatattg caattgatgg taatgaactt
     1501 gacgcactga cgatattagt caatttaacc gataagactg atgatcagaa agacctgctc
     1561 gatcgagcca aatgcaagca aaaacttcga gatgaaaaat ggtgggctag ctgcataaat
     1621 tgcgttaatt acagacaaag ccataaccca aaattcccgg atatacgttc tgaaggcgtg
     1681 attcgaaccc aagccctggg tgaattaccg agtttcctac tctcgtcctc aaaaatccca
     1741 ccataccatt ggtcatatag ccatgattca aaatacgtca acaaaagcgc attcctcacc
     1801 aatgagtttt gttgggatgg tgagatctca tgtttgggtg agcttcttaa agatgcagat
     1861 cacccacttt ggaatacttt gaaaaagtta ggttgttctc aaaaaacttg caaagcaatg
     1921 gcaaaacaac tagctgatat tactctcacg actatcaatg tcacactcgc accaaattac
     1981 ctgactcaaa tatctctccc cgatagtgat acatcctaca tttcactctc tcctgttgca
     2041 tcgctatcga tgcaaagcca ctttcatcag aggctccaag atgaaaatcg gcatagcgcg
     2101 ataacgcggt ttagccgaac taccaacatg ggggtcactg cgatgacatg tggcggcgca
     2161 tttagaatgt taaagtctgg cgctaagttt tccagccccc ctcatcaccg attaaacagt
     2221 aaacgaagtt ggttgacgtc agagcatgtt cagtcattaa aacagtacca gcgcctcaac
     2281 aaaagcctca tacctgaaaa ctctcggatt gcactccgta gaaaatacaa aatcgagctt
     2341 caaaatatgg tcagatcttg gtttgcaatg caagaccata cccttgattc gaacatactt
     2401 atccaacatt tgaatcatga cctatcttac ttaggagcca caaaacgttt tgcatacgat
     2461 ccagcgatga ccaagctctt tactgagctt ttgaaacgag agttatcaaa ttcaatcaat
     2521 aatggtgagc aacacactaa tggatcgttt ttagtcctac cgaatatcag ggtttgtggc
     2581 gcaacagctt taagctcccc ggtaacggtg gggattccat cacttacagc tttctttggc
     2641 ttcgttcacg catttgaacg gaatataaat cgcaccacct catcgtttcg tgttgaatcc
     2701 tttgcgatat gcgtccatca actacatgtc gagaagcgag gtttgacagc agagtttgtg
     2761 gaaaaaggcg acgggactat atccgctccc gcgacccggg atgactggca gtgtgatgtc
     2821 gtatttagcc ttattttgaa caccaacttt gctcaacata ttgaccaaga tacgttagtt
     2881 acatcactac caaagcgatt ggctcggggt tcagcaaaaa ttgcgattga tgactttaaa
     2941 catatcaact cattctcgac attagaaaca gcgatcgaat ctctgccaat agaagctggt
     3001 aggtggttat cactttacgc acagtcaaac aataatctaa gtgatctatt agcagccatg
     3061 acagaggacc atcagctcat ggcaagctgc gtcggttacc acttgttaga agagcccaaa
     3121 gataaaccaa actccctcag aggttacaaa cacgctatcg ccgagtgcat cattggactc
     3181 attaactcaa tcacctttag ctcagagact gatcccaaca caatcttttg gtcgctaaag
     3241 aactatcaaa actacctagt ggtacagcca aggagtatca acgatgaaac taccgacaaa
     3301 tctagcctat gagcgctcta tcgacccatc agatgtctgt ttttttgtcg tctggcccga
     3361 tgatagaaaa acacctttaa cctacaattc tcgtactctg ctcgggcaaa tggaagcggc
     3421 atcattagcc tatgatgtct caggtcaacc aataaaaagt gccaccgctg aggcgttagc
     3481 tcaagggaac cctcatcaag ttgatttctg ccacgttcca tacggtgcga gtcatattga
     3541 atgcagtttc tccgtctcgt tttcttctga actacgtcaa ccatataagt gtaactcaag
     3601 caaagttaaa caaacgctag tgcaattagt cgagctctac gaaacgaaaa tcggctggac
     3661 tgagctagca acccgatatt tgatgaacat ttgcaacggt aaatggctgt ggaaaaatac
     3721 ccgtaaagcc tattgctgga acattgtact tacaccttgg ccttggaacg gggaaaaggt
     3781 tggatttgaa gatattcgta ctaactacac ctcacggcaa gactttaaaa ataataaaaa
     3841 ttggtctgct atagttgaaa tgatcaaaac cgcattttct agtactgatg ggctggcgat
     3901 atttgaagtc agggccacct tgcacttgcc aacgaatgct atggtgcggc caagccaagt
     3961 tttcacagaa aaagaaagtg gcagtaaaag taaatctaaa actcaaaaca gtcgagtttt
     4021 tcagagtaca actattgatg gtgaacgatc gccaatacta ggggccttta aaacgggagc
     4081 agctattgca accattgacg actggtatcc tgaagccact gagccactaa gggtcggacg
     4141 gtttggggtt catcgcgaag atgtcacttg ctaccgtcat ccgtctaccg gaaaagattt
     4201 tttctcgata ttacaacaag cagagcacta tattgaagtg ttgagcgcca acaaaactcc
     4261 cgctcaagaa actatcaacg acatgcactt tttaatggct aacctgatta agggtgggat
     4321 gttccagcat aaaggagact gactgtgaaa tggtattata agacaatcac ctttctgcca
     4381 gagttgtgca acaacgagtc actggctgca aagtgtctcc gcgttctgca tggatttaac
     4441 tatcagtatg agacacgaaa tatcggcgtt tcatttccgc tttggtgtga tgcaacggtt
     4501 ggaaaaaaga tttcatttgt cagcaagaac aagatagaac tcgacttact acttaaacaa
     4561 cactatttcg tccaaatgga acaacttcaa tactttcata tatccaacac tgttctcgtc
     4621 ccagaagatt gtacatacgt ttcctttaga cgctgtcaat ctatagataa gctcacagca
     4681 gcagggctgg caaggaaaat cagacgcctg gagaaacgtg ctctatctag aggcgagcaa
     4741 tttgacccat catcttttgc tcaaaaagag catactgcaa tagcgcacta ccactcactt
     4801 ggggagtcca gcaaacagac gaaccgcaac tttcgactca atatcaggat gctctcggag
     4861 caaccccgtg aggggaactc gatttttagt agctatggct tatcaaattc agaaaactcg
     4921 tttcagcctg tacccttaat ctgacctagg ctgctgccac cgctgagcaa taactagcat
     4981 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
     5041 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     5101 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg
     5161 cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg gtgatctcgc
     5221 ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt
     5281 cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
     5341 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
     5401 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
     5461 agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat
     5521 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
     5581 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
     5641 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
     5701 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
     5761 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
     5821 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
     5881 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
     5941 ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat
     6001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6061 agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg agactgcggc
     6121 gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag gggactaaca
     6181 tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc gccctcctgc
     6241 cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga ggctcaacca
     6301 tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg tttccggcgg
     6361 gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac ctgttccgcc
     6421 tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta tctcggctcg
     6481 gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca gcccgactgc
     6541 tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg taaaacgcca
     6601 ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta aacacgcggt
     6661 tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg gttgctgtgc
     6721 tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc ttcgatcaaa
     6781 ccacctcccc aggtggtttt ttcgaaggtt aagtcagttg gggaactgct taaccttcga
     6841 tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa
     6901 aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat
     6961 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     7021 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     7081 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     7141 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7201 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     7261 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     7321 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     7381 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
     7441 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     7501 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     7561 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     7621 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     7681 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     7741 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     7801 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     7861 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     7921 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     7981 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     8041 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     8101 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     8161 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     8221 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     8281 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa
     8341 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt
     8401 ttgtttaact ttaataagga gatataccat gggtgaactg ccgagtaggt agctgataac
     8461 ggaacgttat cgttcacacg gaccgtggag gcgtgaactg ccgagtaggt
//
