LOCUS       sequence-346341-        8138 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 8138)
  AUTHORS   Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S
  TITLE     Orthogonal CRISPR-associated transposases for parallel and 
            multiplexed chromosomal integration.
  JOURNAL   Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi: 
            10.1093/nar/gkab752.
  PUBMED    34478496
REFERENCE   2  (bases 1 to 8138)
  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: "Nucleic 
            Acids Res. 2021 Sep 3. pii: 6363765. doi: 10.1093/nar/gkab752."
COMMENT     Sequence Label: pQCascade(Ptr)-entry(BsaI)
FEATURES             Location/Qualifiers
     source          1..8138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..56
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7..25
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        263..318
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    269..287
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    299..317
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5305..5323)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5319..5366
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5537..6328)
                     /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(6329..6420)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7497..8123)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     8028..8047
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 gttgacactc tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt
       61 gaactctaga aataattttg tttaacttta aaaggagata taccatgggt gaactgccga
      121 gtaggcagct ggaaatgaga cctctggtct cgtgaactgc cgagtaggca gctgaagttg
      181 gatccgaatt cgagctcggc gcgcctgcag gtcgacaagc ttgcggccgc tctaatctag
      241 acatcattaa ttcctaattt ttgttgacac tctatcattg atagagttat tttaccactc
      301 cctatcagtg atagagaaaa gtgaactcta gaaataattt tgtttaactt taaaggagat
      361 atacatatgt ttttgcaaag acctaaataa ccaacaagaa ggagatatac atatgcactt
      421 tctggtgcag accaagagct acccggacga ggcgctggaa agctatctgc tgcgtctggc
      481 gcgtgataac agctacaacg gttatagcga gctggcggac atcctgtggc agtggctggc
      541 ggaacaagat aacgagctgg aaggtgcgct gccgctggcg ctgagcaagg tggacgttta
      601 ccacgcgcgt caggcgagca gcttccgtat ccgtgcgctg aaactggtgg cgcaactggc
      661 ggacgttaac gcgggtgata ttctggcgct ggcgtggcgt cgtagcaact tcaagtttgg
      721 caacctggcg gcggtgagcc gtaacgagct ggcgatcccg ctggaactgc tgcgtaccga
      781 taacatcccg gtttgcatta aatgcctgag cgagagcagc cacattccgt tttactggca
      841 cctgaagccg tataaagcgt gccacaagca caaaagccag ctgatcaccc gttgcaagga
      901 gtgctacgac ctgattgatt atcgtgcgag cgaggcgttt ctggaatgcg tttgcggttg
      961 caaaatcacc aacagcgaac aactgaacga cgcggatttc aagatcgcga ttgcgctggc
     1021 gagcagcaac agccagaaaa tcgtgggcct gattagctgg ttcgcgaagg tgaaacaact
     1081 ggacgttagc gacgcggatt tcaactgcgc gtttgttgat tacttcaaca cctggccgga
     1141 gagcctgacc accgaactgg acctgctgac caacaacgcg cgtctgaagc agctgaaccc
     1201 gtttaacaag accaaattca gcagcgtgta cggtgacctg atccgtgatg gccaaattgc
     1261 ggcgaccagc aaccgtaaga acaaagttat cgacgagatc attagctatt ttgtggaact
     1321 ggttgatagc aacccgaagg cgaaacaccc gaacattggt gacctgctgc tgtgcacctt
     1381 cgatgcggcg gtgctgctga acaccaccac cgagcaggtt taccgtctgc accaagaagc
     1441 gtttctgaac tgcgcgtata gccagaagaa acacgaacaa ctgcgtgcgg atagccacgt
     1501 gttctatctg cgtcaggtta tcgagctgca gcaagcgttt gcggcggaaa aaccgctgac
     1561 caagaaacaa ttcattgcgc cgtggtaact tatgaacctg caggatgcgc tggcgattga
     1621 gccgctgaag gaaaaaacca ccgcgctgcg taagctgttc gtgccgtaca ccagccacgt
     1681 tgaggtggat ggttttgagg aactggcgct gaccgtgctg atcaacctgg tttataagcg
     1741 tagcgaaatt gacgatctga ccagcgcgcg taccgcgaaa agcgtgctgc gtgacgaggt
     1801 tctgctgagc aagtgcatca acgaagtgaa atggttccac acccacaacc tgaagtaccc
     1861 ggacatccgt gttagccacc aacgtctgat tagcgaggtg gttagcgaag atatcgcggg
     1921 tatttgcagc cgtagcctgc cgctgagctt tggctggagc cacaacagcg cggagatcaa
     1981 ccacgcgaaa ctgttcctga ccagctttaa ctggcagggt gaagtgacct gcctggcgcg
     2041 tctgctgatt aacgaggaac cggtttggat caacctgatt cgtgcgtacg gtttcaccaa
     2101 gaaagcggtt ctggagatca gcggcaagat taaacagcaa ctgccggtgg cggagttccc
     2161 gctggaagtt agcagcttta gcccgcagct gcaaatgccg tttcagcaaa gctatctggt
     2221 ggttaccccg gtggttagcc acgcgatgct ggcgaagatc cagcaactga ccaccgaccg
     2281 taaactgaac ttcgcgctgg ttgagcacag ccgtccggcg aacgttggtg atctggcgag
     2341 cagcgtgggt ggcaacattc gtgttctgcg ttactttccg aagacctata gcaaagcggt
     2401 gaaccgtagc aaagttgcga acaacgatat cgaaaaggcg ttcaaaattc gtgcgctgct
     2461 gagcagccag tttcagcaag cgctgctggt gctggttggc atcaagcagt tcaacaccct
     2521 gcgtcaaaaa cgtctggcgc gtgtggcggc gatccgtcaa gtgcgtgtta gcctgcaact
     2581 gtggctggac aacattctgg aggcgaagaa caacgcgcag aaccaagtgt acccggaatg
     2641 ggttcgtcac tatctggatc aaagcatcac caactgcatt agccagttca gcaacgttct
     2701 gaacgaaagc ctgggtaacc tgagcaagct gaaacgtttt gcgtaccacc cgaacctgat
     2761 gggcctgttc aaagcgcaac tgaactatgt gtttacccac tgcgcggcgg agcaggaaat
     2821 cctgaacgac gagcaaattg tgtacgttca ctgccaggac atgcgtgttt tcgatgcgga
     2881 agcgatggcg aacccgtata tccagggtat gccgagcctg accgcgctga acggcctggc
     2941 gcacaacttc gagcgtaagc tgaaaaactt tattgatccg agcatcaagt gcattggtag
     3001 cgcgatctac attgagaact atcaactgca caccggcaaa ccgctgccgg aaccgagcaa
     3061 gctgaaacag gtggcgggtc gtagccacgt tatccgtagc ggcatcattg acaagccgaa
     3121 atgcgacatt accctggatc tggtgttccg tctgtttgtt ccgaacaccg aactgctgga
     3181 taagctgaac agccaactga ttaagccggc gctgccgagc agctttgcgg gtggcaccat
     3241 gcacccgccg agcctgtacc agaacattga ctggtgccac gtgcacacca agccgagcga
     3301 gctgtttaag aaactgaagg cgaaaagcag caacggtagc tggctgtatc cgagcaagaa
     3361 agtggttaaa agcttcgaac agctgatcga cgcgctgaac agcaacttta acctgcgtcc
     3421 ggcggcgatt ggcctggcgg cgctggagga accggtgaag cgtgatgcgg cgctgcacga
     3481 gtaccactgc tatgcggaac cggttatcgg tctgctggag tgcgtgagca acaccagcgt
     3541 taagtacgcg ggcgcgaaac aattctttca cgacgcgttc tgggtgatgg atgttcagaa
     3601 ggaaagcatg ctgatgaaga aaagcaaatt tgagtatgaa taatgcagct gccgcgtcac
     3661 ctgagctaca cccgtagcct gagcccgagc aaggcggtgt tcttttataa aaccccggag
     3721 agcgacttcg aaccgctgca gatcgagcaa aacaaactgg tgggtcagaa gagcggtttt
     3781 ggcgatgcgt accagaagca aaacgttgcg aaaaacctgg cgccgcagga cctggcgttt
     3841 ggtaacccgc aaaccattga tgtgtgctat gttccgccga ccgtgaacga actgttctgc
     3901 cgttttagcc tgcgtgttga ggcgaactgc atcgaaccgc acgtgtgcga cgatccgaag
     3961 gttatttact ggctgaaacg tttctttgaa acctataaga aacacaacgg tctgaacgaa
     4021 gtggcgaccc gttacgcgaa gaacatcctg atgggcaact ggctgtggcg taaccgtcag
     4081 agcccgaacg ttgacatcga gattctgacc gaacacgcgg cgccgattgt ggttgagggt
     4141 gcgcagaagc tgaaatggca aggcaactgg cagaacaacc aaaccgcgct gctgaccctg
     4201 agcgagagca tccaggaagg tctgagcaac ccgcaaaact actgctatct ggatatcacc
     4261 gcgaagatta aaaacgcgtt cagccaggaa gtgcacccga gccaaaagtt tgtggacaac
     4321 gttgaacagg gtatgagcag caaacagctg gcgtataccc aagtgggcga taagaaagcg
     4381 gcgagcctga acagccagaa ggttggcgcg gcgatccaaa ccattgacga ttggtacgag
     4441 gaaggttata aaccgctgcg tacccatgag tatggtgcgg acaagcaaat cctggtggcg
     4501 caccgtaccc cgaaaagcca cagcgatttt tatagcctgc tgccgcgtat cgcgctgcac
     4561 attaagcaca tggaaaaaca cggtctggag cagagcgaac aaagcaacag catccacttc
     4621 attgcggcgg ttctgattaa gggtggcctg tttcagcgta gcaaaggatg aagcgttact
     4681 atttcaccat cacctacctg ccgcaaagct gcgatgtgag cctgctggcg ggtcgttgca
     4741 tcggcattct gcacggtttc atgagcagcc gtgagatcag caacattggc gtgtgctttc
     4801 cgaaatggaa cgagcagacc atcggtaacg aactggcgtt tgttagcacc aacaagaaac
     4861 aactgaccaa cctgagccag caaagctatt tcgagatgat ggcgcacgac aagctgtttg
     4921 gcctgagcaa aattctggaa gtgccggtta accagagcga agtgatgttc gttcgtaacc
     4981 aaagcgtggc gaaggcgttt gttggtgaaa agcaacgtcg tctgaaacgt gcgaagaaac
     5041 gtgcggaggc gcgtggcgaa gtgtacaacc cggagtataa gttcgaagcg aaagatatcg
     5101 gtcactttca cagcattccg gtgagcagca agggtaacgg ccagagctac gttctgcaca
     5161 tccaaaagaa cgagaacgcg gaaagcatta aaaaccagtt caacaactat ggctttgcga
     5221 ccaaccaaat tttcctgggc accgtgccga gcctgaacac cctgctgtaa ggtaccaccc
     5281 ttaatctgac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     5341 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
     5401 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     5461 gaccctgccc tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg
     5521 aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
     5581 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
     5641 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
     5701 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
     5761 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
     5821 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
     5881 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
     5941 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
     6001 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
     6061 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
     6121 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
     6181 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
     6241 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
     6301 atacttcggc gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt
     6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6421 tagctagctc actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac
     6481 atacaaagtt acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag
     6541 cagggccgcg ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa
     6601 cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac
     6661 gggcgaaacc cgacaggact taaagatccc caccgtttcc ggcgggtcgc tccctcttgc
     6721 gctctcctgt tccgaccctg ccgtttaccg gatacctgtt ccgcctttct cccttacggg
     6781 aagtgtggcg ctttctcata gctcacacac tggtatctcg gctcggtgta ggtcgttcgc
     6841 tccaagctgg gctgtaagca agaactcccc gttcagcccg actgctgcgc cttatccggt
     6901 aactgttcac ttgagtccaa cccggaaaag cacggtaaaa cgccactggc agcagccatt
     6961 ggtaactggg agttcgcaga ggatttgttt agctaaacac gcggttgctc ttgaagtgtg
     7021 cgccaaagtc cggctacact ggaaggacag atttggttgc tgtgctctgc gaaagccagt
     7081 taccacggtt aagcagttcc ccaactgact taaccttcga tcaaaccacc tccccaggtg
     7141 gttttttcga aggttaagtc agttggggaa ctgcttaacc ttcgatcaaa ccacctcccc
     7201 aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag aaaaaaagga tctcaagaag
     7261 atcctttgat cttttctact gaaccgctct agatttcagt gcaatttatc tcttcaaatg
     7321 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     7381 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcctga tgaatcccct
     7441 aatgattttt atcaaaatca ttaaggttac catcacggaa aaaggttatg ctgcttttaa
     7501 gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc aaggccgaat
     7561 aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg taataatggc
     7621 ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg ctcttgatct
     7681 tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa tgcattctct
     7741 agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc atactgtttt
     7801 tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag taaagcacat
     7861 ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc taaagggcaa
     7921 aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa agcccgctta
     7981 ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc gagtttacgg
     8041 gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt aatcacttta
     8101 cttttatcta atctagacat cattaattcc taattttt
//
