LOCUS       sequence-306000-        8493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and CRISPR RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8493)
  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 8493)
  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
FEATURES             Location/Qualifiers
     source          1..8493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    146..170
                     /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(4968..4986)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4982..5029
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5200..5991)
                     /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(5992..6083)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7144..8226)
                     /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     8188..8207
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8227..8304)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        8350..8368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8369..8393
                     /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 gagtaggtag ctgataacgg atccgaattc gagctcggcg cgcctgcagg tcgacaagct
       61 tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
      121 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
      181 atattagtta agtataagaa ggagatatac atatgttttt gcaaagacct aaaccttaca
      241 gcgatgaaag tttagaaagt ttctttatcc gagtggctaa caaaaatggc tacggtgatg
      301 tccatcgctt cctagaagcc actaaacgat tccttcaaga cattgaccat aatggctatc
      361 aaacctttcc gactgatata actcggataa acccatactc agctaaaaac agttccagcg
      421 cacgaactgc gtcattcctg aaacttgcac aattgacatt taatgaaccg ccagagctac
      481 ttgggttggc aattaacaga acaaacatga aatactcgcc gtcaactagc gcggttgtta
      541 gaggtgcaga agtctttcct cgcagtttac tacggacgca ctccatcccc tgctgtcctt
      601 tgtgtctgcg agaaaatggc tacgcctcct acctttggca ctttcagggg tacgaatact
      661 gccacagcca taacgtacct ttaattacca cttgtagctg tggtaaggag tttgactacc
      721 gagtatctgg gttaaagggc atttgctgca aatgcaagga gcctatcacc ttaaccagca
      781 gggagaacgg tcatgaggca gcgtgtactg tttcaaactg gcttgctggc catgaatcta
      841 aacctctgcc aaatcttcct aaaagctacc gatggggttt agttcattgg tggatgggta
      901 ttaaagatag cgagttcgat cacttttcgt tcgttcaatt tttctcaaac tggccaaggt
      961 cattccactc gataatcgaa gatgaagtag agttcaacct tgagcatgct gttgtcagca
     1021 cgtctgaatt acgactaaaa gatcttcttg gtcgattgtt tttcggttca attcggttac
     1081 ctgagcggaa tcttcaacac aatatcatcc ttggtgagct tctctgctat ttagaaaatc
     1141 gtttatggca agacaaggga ttaatcgcca acctcaaaat gaacgcgtta gaggcgactg
     1201 taatgttaaa ttgtagcctc gatcagattg catcaatggt tgaacaacgc atcttgaagc
     1261 caaatcgaaa aagcaagccc aacagccctc ttgatgttac cgattatcta tttcatttcg
     1321 gcgatatttt ctgtctttgg ttagctgagt tccaaagcga tgagtttaac cgttcgtttt
     1381 atgtgtcgag gtggtaaatg caaactctga aagaactaat cgcatccaat cctgacgact
     1441 taacaactga acttaagaga gcatttcgtc cactcacacc gcatattgca attgatggta
     1501 atgaacttga cgcactgacg atattagtca atttaaccga taagactgat gatcagaaag
     1561 acctgctcga tcgagccaaa tgcaagcaaa aacttcgaga tgaaaaatgg tgggctagct
     1621 gcataaattg cgttaattac agacaaagcc ataacccaaa attcccggat atacgttctg
     1681 aaggcgtgat tcgaacccaa gccctgggtg aattaccgag tttcctactc tcgtcctcaa
     1741 aaatcccacc ataccattgg tcatatagcc atgattcaaa atacgtcaac aaaagcgcat
     1801 tcctcaccaa tgagttttgt tgggatggtg agatctcatg tttgggtgag cttcttaaag
     1861 atgcagatca cccactttgg aatactttga aaaagttagg ttgttctcaa aaaacttgca
     1921 aagcaatggc aaaacaacta gctgatatta ctctcacgac tatcaatgtc acactcgcac
     1981 caaattacct gactcaaata tctctccccg atagtgatac atcctacatt tcactctctc
     2041 ctgttgcatc gctatcgatg caaagccact ttcatcagag gctccaagat gaaaatcggc
     2101 atagcgcgat aacgcggttt agccgaacta ccaacatggg ggtcactgcg atgacatgtg
     2161 gcggcgcatt tagaatgtta aagtctggcg ctaagttttc cagcccccct catcaccgat
     2221 taaacagtaa acgaagttgg ttgacgtcag agcatgttca gtcattaaaa cagtaccagc
     2281 gcctcaacaa aagcctcata cctgaaaact ctcggattgc actccgtaga aaatacaaaa
     2341 tcgagcttca aaatatggtc agatcttggt ttgcaatgca agaccatacc cttgattcga
     2401 acatacttat ccaacatttg aatcatgacc tatcttactt aggagccaca aaacgttttg
     2461 catacgatcc agcgatgacc aagctcttta ctgagctttt gaaacgagag ttatcaaatt
     2521 caatcaataa tggtgagcaa cacactaatg gatcgttttt agtcctaccg aatatcaggg
     2581 tttgtggcgc aacagcttta agctccccgg taacggtggg gattccatca cttacagctt
     2641 tctttggctt cgttcacgca tttgaacgga atataaatcg caccacctca tcgtttcgtg
     2701 ttgaatcctt tgcgatatgc gtccatcaac tacatgtcga gaagcgaggt ttgacagcag
     2761 agtttgtgga aaaaggcgac gggactatat ccgctcccgc gacccgggat gactggcagt
     2821 gtgatgtcgt atttagcctt attttgaaca ccaactttgc tcaacatatt gaccaagata
     2881 cgttagttac atcactacca aagcgattgg ctcggggttc agcaaaaatt gcgattgatg
     2941 actttaaaca tatcaactca ttctcgacat tagaaacagc gatcgaatct ctgccaatag
     3001 aagctggtag gtggttatca ctttacgcac agtcaaacaa taatctaagt gatctattag
     3061 cagccatgac agaggaccat cagctcatgg caagctgcgt cggttaccac ttgttagaag
     3121 agcccaaaga taaaccaaac tccctcagag gttacaaaca cgctatcgcc gagtgcatca
     3181 ttggactcat taactcaatc acctttagct cagagactga tcccaacaca atcttttggt
     3241 cgctaaagaa ctatcaaaac tacctagtgg tacagccaag gagtatcaac gatgaaacta
     3301 ccgacaaatc tagcctatga gcgctctatc gacccatcag atgtctgttt ttttgtcgtc
     3361 tggcccgatg atagaaaaac acctttaacc tacaattctc gtactctgct cgggcaaatg
     3421 gaagcggcat cattagccta tgatgtctca ggtcaaccaa taaaaagtgc caccgctgag
     3481 gcgttagctc aagggaaccc tcatcaagtt gatttctgcc acgttccata cggtgcgagt
     3541 catattgaat gcagtttctc cgtctcgttt tcttctgaac tacgtcaacc atataagtgt
     3601 aactcaagca aagttaaaca aacgctagtg caattagtcg agctctacga aacgaaaatc
     3661 ggctggactg agctagcaac ccgatatttg atgaacattt gcaacggtaa atggctgtgg
     3721 aaaaataccc gtaaagccta ttgctggaac attgtactta caccttggcc ttggaacggg
     3781 gaaaaggttg gatttgaaga tattcgtact aactacacct cacggcaaga ctttaaaaat
     3841 aataaaaatt ggtctgctat agttgaaatg atcaaaaccg cattttctag tactgatggg
     3901 ctggcgatat ttgaagtcag ggccaccttg cacttgccaa cgaatgctat ggtgcggcca
     3961 agccaagttt tcacagaaaa agaaagtggc agtaaaagta aatctaaaac tcaaaacagt
     4021 cgagtttttc agagtacaac tattgatggt gaacgatcgc caatactagg ggcctttaaa
     4081 acgggagcag ctattgcaac cattgacgac tggtatcctg aagccactga gccactaagg
     4141 gtcggacggt ttggggttca tcgcgaagat gtcacttgct accgtcatcc gtctaccgga
     4201 aaagattttt tctcgatatt acaacaagca gagcactata ttgaagtgtt gagcgccaac
     4261 aaaactcccg ctcaagaaac tatcaacgac atgcactttt taatggctaa cctgattaag
     4321 ggtgggatgt tccagcataa aggagactga ctgtgaaatg gtattataag acaatcacct
     4381 ttctgccaga gttgtgcaac aacgagtcac tggctgcaaa gtgtctccgc gttctgcatg
     4441 gatttaacta tcagtatgag acacgaaata tcggcgtttc atttccgctt tggtgtgatg
     4501 caacggttgg aaaaaagatt tcatttgtca gcaagaacaa gatagaactc gacttactac
     4561 ttaaacaaca ctatttcgtc caaatggaac aacttcaata ctttcatata tccaacactg
     4621 ttctcgtccc agaagattgt acatacgttt cctttagacg ctgtcaatct atagataagc
     4681 tcacagcagc agggctggca aggaaaatca gacgcctgga gaaacgtgct ctatctagag
     4741 gcgagcaatt tgacccatca tcttttgctc aaaaagagca tactgcaata gcgcactacc
     4801 actcacttgg ggagtccagc aaacagacga accgcaactt tcgactcaat atcaggatgc
     4861 tctcggagca accccgtgag gggaactcga tttttagtag ctatggctta tcaaattcag
     4921 aaaactcgtt tcagcctgta cccttaatct gacctaggct gctgccaccg ctgagcaata
     4981 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
     5041 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     5101 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc
     5161 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
     5221 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     5281 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     5341 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     5401 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     5461 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     5521 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     5581 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     5641 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     5701 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     5761 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     5821 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     5881 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     5941 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct
     6001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6061 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag
     6121 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg
     6181 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc
     6241 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg
     6301 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt
     6361 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct
     6421 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc
     6481 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc
     6541 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta
     6601 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa
     6661 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt
     6721 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt
     6781 cgatcaaacc acctccccag gtggtttttt cgaaggttaa gtcagttggg gaactgctta
     6841 accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa agattacgcg
     6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc tctagatttc
     6961 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     7021 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     7081 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     7141 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     7261 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     7321 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     7381 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     7441 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     7501 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     7561 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     7621 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     7681 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     7741 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     7801 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     7861 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     7921 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     7981 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     8041 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     8101 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     8161 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     8221 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     8281 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     8341 ttaggaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctgtag
     8401 aaataatttt gtttaacttt aataaggaga tataccatgg gtgaactgcc gagtaggtag
     8461 ctgataacga gacctctggt ctcgtgaact gcc
//
