LOCUS       sequence-306003-        8865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-array1. 
            The crRNA-array1 targets 8 different loci in Escherchia coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8865)
  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 8865)
  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-array1
FEATURES             Location/Qualifiers
     source          1..8865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     585..604
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(624..701)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        747..765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    766..790
                     /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        1454..1472
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1473..1497
                     /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(6295..6313)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6309..6356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6527..7318)
                     /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(7319..7410)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7458..8196)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(join(8406..8865,1..623))
                     /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 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
       61 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
      121 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
      181 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
      241 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
      301 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
      361 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
      421 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
      481 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
      541 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
      601 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
      661 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
      721 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
      781 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat accatgggtg
      841 aactgccgag taggtagctg ataacgcctg gcttggcgat gctcgcctga tcgacaagtg
      901 aactgccgag taggtagctg ataacgagaa tattgtggca aaagcgcata aggtgctgtg
      961 aactgccgag taggtagctg ataacgtcag agtggcgtat ccgatgaatc accacaggtg
     1021 aactgccgag taggtagctg ataacgcaga agtgcacggc gtgtgcggcg gcgagcagtg
     1081 aactgccgag taggtagctg ataactcgat ccgtgggtgt ttatgttctc agcaggagtg
     1141 aactgccgag taggtagctg ataacttact tacggcgttg cgcattctca ttgcaccgtg
     1201 aactgccgag taggtagctg ataacgcatt actaagctgg cgcgtgaagc tgtagaagtg
     1261 aactgccgag taggtagctg ataacggggc gtaaaaagaa ggggcgctga ttaacgcgtg
     1321 aactgccgag taggtagctg ataacggatc cgaattcgag ctcggcgcgc ctgcaggtcg
     1381 acaagcttgc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc
     1441 gcataatcga aattaatacg actcactata ggggaattgt gagcggataa caattcccca
     1501 tcttagtata ttagttaagt ataagaagga gatatacata tgtttttgca aagacctaaa
     1561 ccttacagcg atgaaagttt agaaagtttc tttatccgag tggctaacaa aaatggctac
     1621 ggtgatgtcc atcgcttcct agaagccact aaacgattcc ttcaagacat tgaccataat
     1681 ggctatcaaa cctttccgac tgatataact cggataaacc catactcagc taaaaacagt
     1741 tccagcgcac gaactgcgtc attcctgaaa cttgcacaat tgacatttaa tgaaccgcca
     1801 gagctacttg ggttggcaat taacagaaca aacatgaaat actcgccgtc aactagcgcg
     1861 gttgttagag gtgcagaagt ctttcctcgc agtttactac ggacgcactc catcccctgc
     1921 tgtcctttgt gtctgcgaga aaatggctac gcctcctacc tttggcactt tcaggggtac
     1981 gaatactgcc acagccataa cgtaccttta attaccactt gtagctgtgg taaggagttt
     2041 gactaccgag tatctgggtt aaagggcatt tgctgcaaat gcaaggagcc tatcacctta
     2101 accagcaggg agaacggtca tgaggcagcg tgtactgttt caaactggct tgctggccat
     2161 gaatctaaac ctctgccaaa tcttcctaaa agctaccgat ggggtttagt tcattggtgg
     2221 atgggtatta aagatagcga gttcgatcac ttttcgttcg ttcaattttt ctcaaactgg
     2281 ccaaggtcat tccactcgat aatcgaagat gaagtagagt tcaaccttga gcatgctgtt
     2341 gtcagcacgt ctgaattacg actaaaagat cttcttggtc gattgttttt cggttcaatt
     2401 cggttacctg agcggaatct tcaacacaat atcatccttg gtgagcttct ctgctattta
     2461 gaaaatcgtt tatggcaaga caagggatta atcgccaacc tcaaaatgaa cgcgttagag
     2521 gcgactgtaa tgttaaattg tagcctcgat cagattgcat caatggttga acaacgcatc
     2581 ttgaagccaa atcgaaaaag caagcccaac agccctcttg atgttaccga ttatctattt
     2641 catttcggcg atattttctg tctttggtta gctgagttcc aaagcgatga gtttaaccgt
     2701 tcgttttatg tgtcgaggtg gtaaatgcaa actctgaaag aactaatcgc atccaatcct
     2761 gacgacttaa caactgaact taagagagca tttcgtccac tcacaccgca tattgcaatt
     2821 gatggtaatg aacttgacgc actgacgata ttagtcaatt taaccgataa gactgatgat
     2881 cagaaagacc tgctcgatcg agccaaatgc aagcaaaaac ttcgagatga aaaatggtgg
     2941 gctagctgca taaattgcgt taattacaga caaagccata acccaaaatt cccggatata
     3001 cgttctgaag gcgtgattcg aacccaagcc ctgggtgaat taccgagttt cctactctcg
     3061 tcctcaaaaa tcccaccata ccattggtca tatagccatg attcaaaata cgtcaacaaa
     3121 agcgcattcc tcaccaatga gttttgttgg gatggtgaga tctcatgttt gggtgagctt
     3181 cttaaagatg cagatcaccc actttggaat actttgaaaa agttaggttg ttctcaaaaa
     3241 acttgcaaag caatggcaaa acaactagct gatattactc tcacgactat caatgtcaca
     3301 ctcgcaccaa attacctgac tcaaatatct ctccccgata gtgatacatc ctacatttca
     3361 ctctctcctg ttgcatcgct atcgatgcaa agccactttc atcagaggct ccaagatgaa
     3421 aatcggcata gcgcgataac gcggtttagc cgaactacca acatgggggt cactgcgatg
     3481 acatgtggcg gcgcatttag aatgttaaag tctggcgcta agttttccag cccccctcat
     3541 caccgattaa acagtaaacg aagttggttg acgtcagagc atgttcagtc attaaaacag
     3601 taccagcgcc tcaacaaaag cctcatacct gaaaactctc ggattgcact ccgtagaaaa
     3661 tacaaaatcg agcttcaaaa tatggtcaga tcttggtttg caatgcaaga ccataccctt
     3721 gattcgaaca tacttatcca acatttgaat catgacctat cttacttagg agccacaaaa
     3781 cgttttgcat acgatccagc gatgaccaag ctctttactg agcttttgaa acgagagtta
     3841 tcaaattcaa tcaataatgg tgagcaacac actaatggat cgtttttagt cctaccgaat
     3901 atcagggttt gtggcgcaac agctttaagc tccccggtaa cggtggggat tccatcactt
     3961 acagctttct ttggcttcgt tcacgcattt gaacggaata taaatcgcac cacctcatcg
     4021 tttcgtgttg aatcctttgc gatatgcgtc catcaactac atgtcgagaa gcgaggtttg
     4081 acagcagagt ttgtggaaaa aggcgacggg actatatccg ctcccgcgac ccgggatgac
     4141 tggcagtgtg atgtcgtatt tagccttatt ttgaacacca actttgctca acatattgac
     4201 caagatacgt tagttacatc actaccaaag cgattggctc ggggttcagc aaaaattgcg
     4261 attgatgact ttaaacatat caactcattc tcgacattag aaacagcgat cgaatctctg
     4321 ccaatagaag ctggtaggtg gttatcactt tacgcacagt caaacaataa tctaagtgat
     4381 ctattagcag ccatgacaga ggaccatcag ctcatggcaa gctgcgtcgg ttaccacttg
     4441 ttagaagagc ccaaagataa accaaactcc ctcagaggtt acaaacacgc tatcgccgag
     4501 tgcatcattg gactcattaa ctcaatcacc tttagctcag agactgatcc caacacaatc
     4561 ttttggtcgc taaagaacta tcaaaactac ctagtggtac agccaaggag tatcaacgat
     4621 gaaactaccg acaaatctag cctatgagcg ctctatcgac ccatcagatg tctgtttttt
     4681 tgtcgtctgg cccgatgata gaaaaacacc tttaacctac aattctcgta ctctgctcgg
     4741 gcaaatggaa gcggcatcat tagcctatga tgtctcaggt caaccaataa aaagtgccac
     4801 cgctgaggcg ttagctcaag ggaaccctca tcaagttgat ttctgccacg ttccatacgg
     4861 tgcgagtcat attgaatgca gtttctccgt ctcgttttct tctgaactac gtcaaccata
     4921 taagtgtaac tcaagcaaag ttaaacaaac gctagtgcaa ttagtcgagc tctacgaaac
     4981 gaaaatcggc tggactgagc tagcaacccg atatttgatg aacatttgca acggtaaatg
     5041 gctgtggaaa aatacccgta aagcctattg ctggaacatt gtacttacac cttggccttg
     5101 gaacggggaa aaggttggat ttgaagatat tcgtactaac tacacctcac ggcaagactt
     5161 taaaaataat aaaaattggt ctgctatagt tgaaatgatc aaaaccgcat tttctagtac
     5221 tgatgggctg gcgatatttg aagtcagggc caccttgcac ttgccaacga atgctatggt
     5281 gcggccaagc caagttttca cagaaaaaga aagtggcagt aaaagtaaat ctaaaactca
     5341 aaacagtcga gtttttcaga gtacaactat tgatggtgaa cgatcgccaa tactaggggc
     5401 ctttaaaacg ggagcagcta ttgcaaccat tgacgactgg tatcctgaag ccactgagcc
     5461 actaagggtc ggacggtttg gggttcatcg cgaagatgtc acttgctacc gtcatccgtc
     5521 taccggaaaa gattttttct cgatattaca acaagcagag cactatattg aagtgttgag
     5581 cgccaacaaa actcccgctc aagaaactat caacgacatg cactttttaa tggctaacct
     5641 gattaagggt gggatgttcc agcataaagg agactgactg tgaaatggta ttataagaca
     5701 atcacctttc tgccagagtt gtgcaacaac gagtcactgg ctgcaaagtg tctccgcgtt
     5761 ctgcatggat ttaactatca gtatgagaca cgaaatatcg gcgtttcatt tccgctttgg
     5821 tgtgatgcaa cggttggaaa aaagatttca tttgtcagca agaacaagat agaactcgac
     5881 ttactactta aacaacacta tttcgtccaa atggaacaac ttcaatactt tcatatatcc
     5941 aacactgttc tcgtcccaga agattgtaca tacgtttcct ttagacgctg tcaatctata
     6001 gataagctca cagcagcagg gctggcaagg aaaatcagac gcctggagaa acgtgctcta
     6061 tctagaggcg agcaatttga cccatcatct tttgctcaaa aagagcatac tgcaatagcg
     6121 cactaccact cacttgggga gtccagcaaa cagacgaacc gcaactttcg actcaatatc
     6181 aggatgctct cggagcaacc ccgtgagggg aactcgattt ttagtagcta tggcttatca
     6241 aattcagaaa actcgtttca gcctgtaccc ttaatctgac ctaggctgct gccaccgctg
     6301 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     6361 aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
     6421 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat
     6481 cgtggccgga tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta
     6541 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     6601 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     6661 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     6721 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag
     6781 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     6841 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     6901 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     6961 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     7021 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     7081 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     7141 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc
     7201 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     7261 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac
     7321 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7381 tatttgaatg tatttagaaa aataaacaaa tagctagctc actcggtcgc tacgctccgg
     7441 gcgtgagact gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata
     7501 agcaggggac taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag
     7561 gctccgccct cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc
     7621 cgtgaggctc aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc
     7681 caccgtttcc ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg
     7741 gatacctgtt ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac
     7801 tggtatctcg gctcggtgta ggtcgttcgc tccaagctgg gctgtaagca agaactcccc
     7861 gttcagcccg actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag
     7921 cacggtaaaa cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt
     7981 agctaaacac gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag
     8041 atttggttgc tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact
     8101 taaccttcga tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg
     8161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt
     8221 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     8281 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     8341 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     8401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     8461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     8521 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     8581 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     8641 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     8701 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     8761 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     8821 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttga
//
