LOCUS       sequence-347616-        9450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CRISPR-associated transposases.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9450)
  AUTHORS   Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, 
            Yang S
  TITLE     Programming Cells by Multicopy Chromosomal Integration Using 
            CRISPR-Associated Transposases.
  JOURNAL   CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018.
  PUBMED    34152213
REFERENCE   2  (bases 1 to 9450)
  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"; doi: 
            "10.1089/crispr.2021"; journalName: "CRISPR J"; date: "2021-06"; 
            volume: "4"; issue: "3"; pages: "350-359"
COMMENT     Sequence Label: pRhaQCas-BsaI
FEATURES             Location/Qualifiers
     source          1..9450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(135..983)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(1057..1893)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        2039..2157
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     promoter        2453..2571
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     primer_bind     complement(7344..7362)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7358..7405
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(7576..8367)
                     /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(8368..8459)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggctctcaag ggcatcggtt cttatggctc ttgtatctat cagtgaagca tcaagactaa
       61 caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg tccataccca
      121 tggggcatgg cgaattaatc tttctgcgaa ttgagatgac gccactggct gggcgtcatc
      181 ccggtttccc gggtaaacac caccgaaaaa tagttactat cttcaaagcc acattcggtc
      241 gaaatatcac tgattaacag gcggctatgc tggagaagat attgcgcatg acacactctg
      301 acctgtcgca gatattgatt gatggtcatt ccagtctgct ggcgaaattg ctgacgcaaa
      361 acgcgctcac tgcacgatgc ctcatcacaa aatttatcca gcgcaaaggg acttttcagg
      421 ctagccgcca gccgggtaat cagcttatcc agcaacgttt cgctggatgt tggcggcaac
      481 gaatcactgg tgtaacgatg gcgattcagc aacatcacca actgcccgaa cagcaactca
      541 gccatttcgt tagcaaacgg cacatgctga ctactttcat gctcaagctg accgataacc
      601 tgccgcgcct gcgccatccc catgctacct aagcgccagt gtggttgccc tgcgctggcg
      661 ttaaatcccg gaatcgcccc ctgccagtca agattcagct tcagacgctc cgggcaataa
      721 ataatattct gcaaaaccag atcgttaacg gaagcgtagg agtgtttatc gtcagcatga
      781 atgtaaaaga gatcgccacg ggtaatgcga taagggcgat cgttgagtac atgcaggcca
      841 ttaccgcgcc agacaatcac cagctcacaa aaatcatgtg tatgttcagc aaagacatct
      901 tgcggataac ggtcagccac agcgactgcc tgctggtcgc tggcaaaaaa atcatctttg
      961 agaagtttta actgatgcgc caccgtggct acctcggcca gagaacgaag ttgattattc
     1021 gcaatatggc gtacaaatac gttgagaaga ttcgcgttat tgcagaaagc catcccgtcc
     1081 ctggcgaata tcacgcggtg accagttaaa ctctcggcga aaaagcgtcg aaaagtggtt
     1141 actgtcgctg aatccacagc gataggcgat gtcagtaacg ctggcctcgc tgtggcgtag
     1201 cagatgtcgg gctttcatca gtcgcaggcg gttcaggtat cgctgaggcg tcagtcccgt
     1261 ttgctgctta agctgccgat gtagcgtacg cagtgaaaga gaaaattgat ccgccacggc
     1321 atcccaattc acctcatcgg caaaatggtc ctccagccag gccagaagca agttgagacg
     1381 tgatgcgctg ttttccaggt tctcctgcaa actgctttta cgcagcaaga gcagtaattg
     1441 cataaacaag atctcgcgac tggcggtcga gggtaaatca ttttcccctt cctgctgttc
     1501 catctgtgca accagctgtc gcacctgctg caatacgctg tggttaacgc gccagtgaga
     1561 cggatactgc ccatccagct cttgtggcag caactgattc agcccggcga gaaactgaaa
     1621 tcgatccggc gagcgataca gcacattggt cagacacaga ttatcggtat gttcatacag
     1681 atgccgatca tgatcgcgta cgaaacagac cgtgccaccg gtgatggtat agggctgccc
     1741 attaaacaca tgaatacccg tgccatgttc gacaatcaca atttcatgaa aatcatgatg
     1801 atgttcagga aaatccgcct gcgggagccg gggttctatc gccacggacg cgttaccaga
     1861 cggaaaaaaa tccacactat gtaatacggt catactggcc tcctgatgtc gtcaacacgg
     1921 cgaaatagta atcacgaggt caggttctta ccttaaattt tcgacggaaa accacgtaaa
     1981 aaacgtcgat ttttcaagat acagcgtgaa ttttcaggaa atgcggtgag catcacatca
     2041 ccacaattca gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc
     2101 attttcctgt cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtaaag
     2161 gagatatacc atgggtgaac tgccgagtag gtagctgata acgagacctc tggtctcgtg
     2221 aactgccgag taggtagctg ataacggatc cgaattcgag ctcggcgcgc ctgcaggtcg
     2281 acaagcttgc ggccgcaata cggtcatact ggcctcctga tgtcgtcaac acggcgaaat
     2341 agtaatcacg aggtcaggtt cttaccttaa attttcgacg gaaaaccacg taaaaaacgt
     2401 cgatttttca agatacagcg tgaattttca ggaaatgcgg tgagcatcac atcaccacaa
     2461 ttcagcaaat tgtgaacatc atcacgttca tctttccctg gttgccaatg gcccattttc
     2521 ctgtcagtaa cgagaaggtc gcgaattcag gcgcttttta gactggtcgt aataaaggag
     2581 atatacatat gtttttgcaa agacctaaac cttacagcga tgaaagttta gaaagtttct
     2641 ttatccgagt ggctaacaaa aatggctacg gtgatgtcca tcgcttccta gaagccacta
     2701 aacgattcct tcaagacatt gaccataatg gctatcaaac ctttccgact gatataactc
     2761 ggataaaccc atactcagct aaaaacagtt ccagcgcacg aactgcgtca ttcctgaaac
     2821 ttgcacaatt gacatttaat gaaccgccag agctacttgg gttggcaatt aacagaacaa
     2881 acatgaaata ctcgccgtca actagcgcgg ttgttagagg tgcagaagtc tttcctcgca
     2941 gtttactacg gacgcactcc atcccctgct gtcctttgtg tctgcgagaa aatggctacg
     3001 cctcctacct ttggcacttt caggggtacg aatactgcca cagccataac gtacctttaa
     3061 ttaccacttg tagctgtggt aaggagtttg actaccgagt atctgggtta aagggcattt
     3121 gctgcaaatg caaggagcct atcaccttaa ccagcaggga gaacggtcat gaggcagcgt
     3181 gtactgtttc aaactggctt gctggccatg aatctaaacc tctgccaaat cttcctaaaa
     3241 gctaccgatg gggtttagtt cattggtgga tgggtattaa agatagcgag ttcgatcact
     3301 tttcgttcgt tcaatttttc tcaaactggc caaggtcatt ccactcgata atcgaagatg
     3361 aagtagagtt caaccttgag catgctgttg tcagcacgtc tgaattacga ctaaaagatc
     3421 ttcttggtcg attgtttttc ggttcaattc ggttacctga gcggaatctt caacacaata
     3481 tcatccttgg tgagcttctc tgctatttag aaaatcgttt atggcaagac aagggattaa
     3541 tcgccaacct caaaatgaac gcgttagagg cgactgtaat gttaaattgt agcctcgatc
     3601 agattgcatc aatggttgaa caacgcatct tgaagccaaa tcgaaaaagc aagcccaaca
     3661 gccctcttga tgttaccgat tatctatttc atttcggcga tattttctgt ctttggttag
     3721 ctgagttcca aagcgatgag tttaaccgtt cgttttatgt gtcgaggtgg taaatgcaaa
     3781 ctctgaaaga actaatcgca tccaatcctg acgacttaac aactgaactt aagagagcat
     3841 ttcgtccact cacaccgcat attgcaattg atggtaatga acttgacgca ctgacgatat
     3901 tagtcaattt aaccgataag actgatgatc agaaagacct gctcgatcga gccaaatgca
     3961 agcaaaaact tcgagatgaa aaatggtggg ctagctgcat aaattgcgtt aattacagac
     4021 aaagccataa cccaaaattc ccggatatac gttctgaagg cgtgattcga acccaagccc
     4081 tgggtgaatt accgagtttc ctactctcgt cctcaaaaat cccaccatac cattggtcat
     4141 atagccatga ttcaaaatac gtcaacaaaa gcgcattcct caccaatgag ttttgttggg
     4201 atggtgagat ctcatgtttg ggtgagcttc ttaaagatgc agatcaccca ctttggaata
     4261 ctttgaaaaa gttaggttgt tctcaaaaaa cttgcaaagc aatggcaaaa caactagctg
     4321 atattactct cacgactatc aatgtcacac tcgcaccaaa ttacctgact caaatatctc
     4381 tccccgatag tgatacatcc tacatttcac tctctcctgt tgcatcgcta tcgatgcaaa
     4441 gccactttca tcagaggctc caagatgaaa atcggcatag cgcgataacg cggtttagcc
     4501 gaactaccaa catgggggtc actgcgatga catgtggcgg cgcatttaga atgttaaagt
     4561 ctggcgctaa gttttccagc ccccctcatc accgattaaa cagtaaacga agttggttga
     4621 cgtcagagca tgttcagtca ttaaaacagt accagcgcct caacaaaagc ctcatacctg
     4681 aaaactctcg gattgcactc cgtagaaaat acaaaatcga gcttcaaaat atggtcagat
     4741 cttggtttgc aatgcaagac catacccttg attcgaacat acttatccaa catttgaatc
     4801 atgacctatc ttacttagga gccacaaaac gttttgcata cgatccagcg atgaccaagc
     4861 tctttactga gcttttgaaa cgagagttat caaattcaat caataatggt gagcaacaca
     4921 ctaatggatc gtttttagtc ctaccgaata tcagggtttg tggcgcaaca gctttaagct
     4981 ccccggtaac ggtggggatt ccatcactta cagctttctt tggcttcgtt cacgcatttg
     5041 aacggaatat aaatcgcacc acctcatcgt ttcgtgttga atcctttgcg atatgcgtcc
     5101 atcaactaca tgtcgagaag cgaggtttga cagcagagtt tgtggaaaaa ggcgacggga
     5161 ctatatccgc tcccgcgacc cgggatgact ggcagtgtga tgtcgtattt agccttattt
     5221 tgaacaccaa ctttgctcaa catattgacc aagatacgtt agttacatca ctaccaaagc
     5281 gattggctcg gggttcagca aaaattgcga ttgatgactt taaacatatc aactcattct
     5341 cgacattaga aacagcgatc gaatctctgc caatagaagc tggtaggtgg ttatcacttt
     5401 acgcacagtc aaacaataat ctaagtgatc tattagcagc catgacagag gaccatcagc
     5461 tcatggcaag ctgcgtcggt taccacttgt tagaagagcc caaagataaa ccaaactccc
     5521 tcagaggtta caaacacgct atcgccgagt gcatcattgg actcattaac tcaatcacct
     5581 ttagctcaga gactgatccc aacacaatct tttggtcgct aaagaactat caaaactacc
     5641 tagtggtaca gccaaggagt atcaacgatg aaactaccga caaatctagc ctatgagcgc
     5701 tctatcgacc catcagatgt ctgttttttt gtcgtctggc ccgatgatag aaaaacacct
     5761 ttaacctaca attctcgtac tctgctcggg caaatggaag cggcatcatt agcctatgat
     5821 gtctcaggtc aaccaataaa aagtgccacc gctgaggcgt tagctcaagg gaaccctcat
     5881 caagttgatt tctgccacgt tccatacggt gcgagtcata ttgaatgcag tttctccgtc
     5941 tcgttttctt ctgaactacg tcaaccatat aagtgtaact caagcaaagt taaacaaacg
     6001 ctagtgcaat tagtcgagct ctacgaaacg aaaatcggct ggactgagct agcaacccga
     6061 tatttgatga acatttgcaa cggtaaatgg ctgtggaaaa atacccgtaa agcctattgc
     6121 tggaacattg tacttacacc ttggccttgg aacggggaaa aggttggatt tgaagatatt
     6181 cgtactaact acacctcacg gcaagacttt aaaaataata aaaattggtc tgctatagtt
     6241 gaaatgatca aaaccgcatt ttctagtact gatgggctgg cgatatttga agtcagggcc
     6301 accttgcact tgccaacgaa tgctatggtg cggccaagcc aagttttcac agaaaaagaa
     6361 agtggcagta aaagtaaatc taaaactcaa aacagtcgag tttttcagag tacaactatt
     6421 gatggtgaac gatcgccaat actaggggcc tttaaaacgg gagcagctat tgcaaccatt
     6481 gacgactggt atcctgaagc cactgagcca ctaagggtcg gacggtttgg ggttcatcgc
     6541 gaagatgtca cttgctaccg tcatccgtct accggaaaag attttttctc gatattacaa
     6601 caagcagagc actatattga agtgttgagc gccaacaaaa ctcccgctca agaaactatc
     6661 aacgacatgc actttttaat ggctaacctg attaagggtg ggatgttcca gcataaagga
     6721 gactgactgt gaaatggtat tataagacaa tcacctttct gccagagttg tgcaacaacg
     6781 agtcactggc tgcaaagtgt ctccgcgttc tgcatggatt taactatcag tatgagacac
     6841 gaaatatcgg cgtttcattt ccgctttggt gtgatgcaac ggttggaaaa aagatttcat
     6901 ttgtcagcaa gaacaagata gaactcgact tactacttaa acaacactat ttcgtccaaa
     6961 tggaacaact tcaatacttt catatatcca acactgttct cgtcccagaa gattgtacat
     7021 acgtttcctt tagacgctgt caatctatag ataagctcac agcagcaggg ctggcaagga
     7081 aaatcagacg cctggagaaa cgtgctctat ctagaggcga gcaatttgac ccatcatctt
     7141 ttgctcaaaa agagcatact gcaatagcgc actaccactc acttggggag tccagcaaac
     7201 agacgaaccg caactttcga ctcaatatca ggatgctctc ggagcaaccc cgtgagggga
     7261 actcgatttt tagtagctat ggcttatcaa attcagaaaa ctcgtttcag cctgtaccct
     7321 taatctgacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     7381 taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac
     7441 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
     7501 accctgccct gaaccgacga ccgggtcatc gtggccggat cttgcggccc ctcggcttga
     7561 acgaattgtt agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtggac
     7621 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc
     7681 ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
     7741 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
     7801 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
     7861 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
     7921 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
     7981 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
     8041 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
     8101 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
     8161 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
     8221 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
     8281 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
     8341 tacttcggcg atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta
     8401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8461 agctagctca ctcggtcgct acgctccggg cgtgagactg cggcgggcgc tgcggacaca
     8521 tacaaagtta cccacagatt ccgtggataa gcaggggact aacatgtgag gcaaaacagc
     8581 agggccgcgc cggtggcgtt tttccatagg ctccgccctc ctgccagagt tcacataaac
     8641 agacgctttt ccggtgcatc tgtgggagcc gtgaggctca accatgaatc tgacagtacg
     8701 ggcgaaaccc gacaggactt aaagatcccc accgtttccg gcgggtcgct ccctcttgcg
     8761 ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc cgcctttctc ccttacggga
     8821 agtgtggcgc tttctcatag ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct
     8881 ccaagctggg ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc ttatccggta
     8941 actgttcact tgagtccaac ccggaaaagc acggtaaaac gccactggca gcagccattg
     9001 gtaactggga gttcgcagag gatttgttta gctaaacacg cggttgctct tgaagtgtgc
     9061 gccaaagtcc ggctacactg gaaggacaga tttggttgct gtgctctgcg aaagccagtt
     9121 accacggtta agcagttccc caactgactt aaccttcgat caaaccacct ccccaggtgg
     9181 ttttttcgaa ggttaagtca gttggggaac tgcttaacct tcgatcaaac cacctcccca
     9241 ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga
     9301 tcctttgatc ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt
     9361 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     9421 gcccaccgga aggagctgac tgggttggaa
//
