LOCUS       sequence-305995-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS1. The 
            crRNA-IS1 targets IS1 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-IS1
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        147..165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    166..190
                     /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(4988..5006)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5002..5049
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5220..6011)
                     /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(6012..6103)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7164..8246)
                     /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     8208..8227
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8247..8324)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        8370..8388
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8389..8413
                     /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 gctgtcgttc gtgaactgcc gagtaggtag ctgataacgg atccgaattc gagctcggcg
       61 cgcctgcagg tcgacaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
      121 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
      181 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatgttttt
      241 gcaaagacct aaaccttaca gcgatgaaag tttagaaagt ttctttatcc gagtggctaa
      301 caaaaatggc tacggtgatg tccatcgctt cctagaagcc actaaacgat tccttcaaga
      361 cattgaccat aatggctatc aaacctttcc gactgatata actcggataa acccatactc
      421 agctaaaaac agttccagcg cacgaactgc gtcattcctg aaacttgcac aattgacatt
      481 taatgaaccg ccagagctac ttgggttggc aattaacaga acaaacatga aatactcgcc
      541 gtcaactagc gcggttgtta gaggtgcaga agtctttcct cgcagtttac tacggacgca
      601 ctccatcccc tgctgtcctt tgtgtctgcg agaaaatggc tacgcctcct acctttggca
      661 ctttcagggg tacgaatact gccacagcca taacgtacct ttaattacca cttgtagctg
      721 tggtaaggag tttgactacc gagtatctgg gttaaagggc atttgctgca aatgcaagga
      781 gcctatcacc ttaaccagca gggagaacgg tcatgaggca gcgtgtactg tttcaaactg
      841 gcttgctggc catgaatcta aacctctgcc aaatcttcct aaaagctacc gatggggttt
      901 agttcattgg tggatgggta ttaaagatag cgagttcgat cacttttcgt tcgttcaatt
      961 tttctcaaac tggccaaggt cattccactc gataatcgaa gatgaagtag agttcaacct
     1021 tgagcatgct gttgtcagca cgtctgaatt acgactaaaa gatcttcttg gtcgattgtt
     1081 tttcggttca attcggttac ctgagcggaa tcttcaacac aatatcatcc ttggtgagct
     1141 tctctgctat ttagaaaatc gtttatggca agacaaggga ttaatcgcca acctcaaaat
     1201 gaacgcgtta gaggcgactg taatgttaaa ttgtagcctc gatcagattg catcaatggt
     1261 tgaacaacgc atcttgaagc caaatcgaaa aagcaagccc aacagccctc ttgatgttac
     1321 cgattatcta tttcatttcg gcgatatttt ctgtctttgg ttagctgagt tccaaagcga
     1381 tgagtttaac cgttcgtttt atgtgtcgag gtggtaaatg caaactctga aagaactaat
     1441 cgcatccaat cctgacgact taacaactga acttaagaga gcatttcgtc cactcacacc
     1501 gcatattgca attgatggta atgaacttga cgcactgacg atattagtca atttaaccga
     1561 taagactgat gatcagaaag acctgctcga tcgagccaaa tgcaagcaaa aacttcgaga
     1621 tgaaaaatgg tgggctagct gcataaattg cgttaattac agacaaagcc ataacccaaa
     1681 attcccggat atacgttctg aaggcgtgat tcgaacccaa gccctgggtg aattaccgag
     1741 tttcctactc tcgtcctcaa aaatcccacc ataccattgg tcatatagcc atgattcaaa
     1801 atacgtcaac aaaagcgcat tcctcaccaa tgagttttgt tgggatggtg agatctcatg
     1861 tttgggtgag cttcttaaag atgcagatca cccactttgg aatactttga aaaagttagg
     1921 ttgttctcaa aaaacttgca aagcaatggc aaaacaacta gctgatatta ctctcacgac
     1981 tatcaatgtc acactcgcac caaattacct gactcaaata tctctccccg atagtgatac
     2041 atcctacatt tcactctctc ctgttgcatc gctatcgatg caaagccact ttcatcagag
     2101 gctccaagat gaaaatcggc atagcgcgat aacgcggttt agccgaacta ccaacatggg
     2161 ggtcactgcg atgacatgtg gcggcgcatt tagaatgtta aagtctggcg ctaagttttc
     2221 cagcccccct catcaccgat taaacagtaa acgaagttgg ttgacgtcag agcatgttca
     2281 gtcattaaaa cagtaccagc gcctcaacaa aagcctcata cctgaaaact ctcggattgc
     2341 actccgtaga aaatacaaaa tcgagcttca aaatatggtc agatcttggt ttgcaatgca
     2401 agaccatacc cttgattcga acatacttat ccaacatttg aatcatgacc tatcttactt
     2461 aggagccaca aaacgttttg catacgatcc agcgatgacc aagctcttta ctgagctttt
     2521 gaaacgagag ttatcaaatt caatcaataa tggtgagcaa cacactaatg gatcgttttt
     2581 agtcctaccg aatatcaggg tttgtggcgc aacagcttta agctccccgg taacggtggg
     2641 gattccatca cttacagctt tctttggctt cgttcacgca tttgaacgga atataaatcg
     2701 caccacctca tcgtttcgtg ttgaatcctt tgcgatatgc gtccatcaac tacatgtcga
     2761 gaagcgaggt ttgacagcag agtttgtgga aaaaggcgac gggactatat ccgctcccgc
     2821 gacccgggat gactggcagt gtgatgtcgt atttagcctt attttgaaca ccaactttgc
     2881 tcaacatatt gaccaagata cgttagttac atcactacca aagcgattgg ctcggggttc
     2941 agcaaaaatt gcgattgatg actttaaaca tatcaactca ttctcgacat tagaaacagc
     3001 gatcgaatct ctgccaatag aagctggtag gtggttatca ctttacgcac agtcaaacaa
     3061 taatctaagt gatctattag cagccatgac agaggaccat cagctcatgg caagctgcgt
     3121 cggttaccac ttgttagaag agcccaaaga taaaccaaac tccctcagag gttacaaaca
     3181 cgctatcgcc gagtgcatca ttggactcat taactcaatc acctttagct cagagactga
     3241 tcccaacaca atcttttggt cgctaaagaa ctatcaaaac tacctagtgg tacagccaag
     3301 gagtatcaac gatgaaacta ccgacaaatc tagcctatga gcgctctatc gacccatcag
     3361 atgtctgttt ttttgtcgtc tggcccgatg atagaaaaac acctttaacc tacaattctc
     3421 gtactctgct cgggcaaatg gaagcggcat cattagccta tgatgtctca ggtcaaccaa
     3481 taaaaagtgc caccgctgag gcgttagctc aagggaaccc tcatcaagtt gatttctgcc
     3541 acgttccata cggtgcgagt catattgaat gcagtttctc cgtctcgttt tcttctgaac
     3601 tacgtcaacc atataagtgt aactcaagca aagttaaaca aacgctagtg caattagtcg
     3661 agctctacga aacgaaaatc ggctggactg agctagcaac ccgatatttg atgaacattt
     3721 gcaacggtaa atggctgtgg aaaaataccc gtaaagccta ttgctggaac attgtactta
     3781 caccttggcc ttggaacggg gaaaaggttg gatttgaaga tattcgtact aactacacct
     3841 cacggcaaga ctttaaaaat aataaaaatt ggtctgctat agttgaaatg atcaaaaccg
     3901 cattttctag tactgatggg ctggcgatat ttgaagtcag ggccaccttg cacttgccaa
     3961 cgaatgctat ggtgcggcca agccaagttt tcacagaaaa agaaagtggc agtaaaagta
     4021 aatctaaaac tcaaaacagt cgagtttttc agagtacaac tattgatggt gaacgatcgc
     4081 caatactagg ggcctttaaa acgggagcag ctattgcaac cattgacgac tggtatcctg
     4141 aagccactga gccactaagg gtcggacggt ttggggttca tcgcgaagat gtcacttgct
     4201 accgtcatcc gtctaccgga aaagattttt tctcgatatt acaacaagca gagcactata
     4261 ttgaagtgtt gagcgccaac aaaactcccg ctcaagaaac tatcaacgac atgcactttt
     4321 taatggctaa cctgattaag ggtgggatgt tccagcataa aggagactga ctgtgaaatg
     4381 gtattataag acaatcacct ttctgccaga gttgtgcaac aacgagtcac tggctgcaaa
     4441 gtgtctccgc gttctgcatg gatttaacta tcagtatgag acacgaaata tcggcgtttc
     4501 atttccgctt tggtgtgatg caacggttgg aaaaaagatt tcatttgtca gcaagaacaa
     4561 gatagaactc gacttactac ttaaacaaca ctatttcgtc caaatggaac aacttcaata
     4621 ctttcatata tccaacactg ttctcgtccc agaagattgt acatacgttt cctttagacg
     4681 ctgtcaatct atagataagc tcacagcagc agggctggca aggaaaatca gacgcctgga
     4741 gaaacgtgct ctatctagag gcgagcaatt tgacccatca tcttttgctc aaaaagagca
     4801 tactgcaata gcgcactacc actcacttgg ggagtccagc aaacagacga accgcaactt
     4861 tcgactcaat atcaggatgc tctcggagca accccgtgag gggaactcga tttttagtag
     4921 ctatggctta tcaaattcag aaaactcgtt tcagcctgta cccttaatct gacctaggct
     4981 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     5041 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     5101 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     5161 acgaccgggt catcgtggcc ggatcttgcg gcccctcggc ttgaacgaat tgttagacat
     5221 tatttgccga ctaccttggt gatctcgcct ttcacgtagt ggacaaattc ttccaactga
     5281 tctgcgcgcg aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt
     5341 atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc
     5401 ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt
     5461 cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc
     5521 tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag
     5581 gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct
     5641 ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc
     5701 ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg
     5761 cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct
     5821 cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt
     5881 ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg
     5941 agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc
     6001 gcttccctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6061 atgagcggat acatatttga atgtatttag aaaaataaac aaatagctag ctcactcggt
     6121 cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga cacatacaaa gttacccaca
     6181 gattccgtgg ataagcaggg gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg
     6241 cgtttttcca taggctccgc cctcctgcca gagttcacat aaacagacgc ttttccggtg
     6301 catctgtggg agccgtgagg ctcaaccatg aatctgacag tacgggcgaa acccgacagg
     6361 acttaaagat ccccaccgtt tccggcgggt cgctccctct tgcgctctcc tgttccgacc
     6421 ctgccgttta ccggatacct gttccgcctt tctcccttac gggaagtgtg gcgctttctc
     6481 atagctcaca cactggtatc tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa
     6541 gcaagaactc cccgttcagc ccgactgctg cgccttatcc ggtaactgtt cacttgagtc
     6601 caacccggaa aagcacggta aaacgccact ggcagcagcc attggtaact gggagttcgc
     6661 agaggatttg tttagctaaa cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac
     6721 actggaagga cagatttggt tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt
     6781 tccccaactg acttaacctt cgatcaaacc acctccccag gtggtttttt cgaaggttaa
     6841 gtcagttggg gaactgctta accttcgatc aaaccacctc cccaggtggt tttttcgttt
     6901 acagggcaaa agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6961 actgaaccgc tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     7021 cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc
     7081 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     7141 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     7261 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     7321 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     7381 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     7441 gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     7501 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     7561 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     7621 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     7681 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     7741 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     7801 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     7861 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     7921 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     7981 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     8041 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     8101 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     8161 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     8221 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     8281 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     8341 tcccttatgc gactcctgca ttaggaaatt aatacgactc actatagggg aattgtgagc
     8401 ggataacaat tcccctgtag aaataatttt gtttaacttt aataaggaga tataccatgg
     8461 gtgaactgcc gagtaggtag ctgataactg gcacggctgg gacggaagtc
//
