LOCUS       sequence-330751-        8570 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 8570)
  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 8570)
  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: pTetQCas-8+IS186
FEATURES             Location/Qualifiers
     source          1..8570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..74
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    25..43
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    55..73
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        831..886
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    837..855
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    867..885
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5690..5708)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5704..5751
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5922..6713)
                     /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(6714..6805)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7853..7870
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7854..7870
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(7947..8570)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     8478..8497
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 cattaattcc taatttttgt tgacactcta tcattgatag agttatttta ccactcccta
       61 tcagtgatag agaaaagtga actctagaaa taattttgtt taactttaaa ggagatatac
      121 catgggtgaa ctgccgagta ggtagctgat aacgaggtct cgataactgg atgctttgcg
      181 tgcaaaggaa cctgaactcg tgaactgccg agtaggtagc tgataacgcc tggcttggcg
      241 atgctcgcct gatcgacaag tgaactgccg agtaggtagc tgataacgag aatattgtgg
      301 caaaagcgca taaggtgctg tgaactgccg agtaggtagc tgataacgtc agagtggcgt
      361 atccgatgaa tcaccacagg tgaactgccg agtaggtagc tgataacgca gaagtgcacg
      421 gcgtgtgcgg cggcgagcag tgaactgccg agtaggtagc tgataactcg atccgtgggt
      481 gtttatgttc tcagcaggag tgaactgccg agtaggtagc tgataactta cttacggcgt
      541 tgcgcattct cattgcaccg tgaactgccg agtaggtagc tgataacgca ttactaagct
      601 ggcgcgtgaa gctgtagaag tgaactgccg agtaggtagc tgataacggg gcgtaaaaag
      661 aaggggcgct gattaacgcg tgaacgagac ctcgtgaact gccgagtagg tagctgataa
      721 cgtttaaact agggataaca gggtaatgga tccgaattcg agctcggcgc gcctgcaggt
      781 cgacaagctt gcggccgctc taatctagac atcattaatt cctaattttt gttgacactc
      841 tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt gaactctaga
      901 aataattttg tttaacttta aaggagatat acatatgttt ttgcaaagac ctaaacctta
      961 cagcgatgaa agtttagaaa gtttctttat ccgagtggct aacaaaaatg gctacggtga
     1021 tgtccatcgc ttcctagaag ccactaaacg attccttcaa gacattgacc ataatggcta
     1081 tcaaaccttt ccgactgata taactcggat aaacccatac tcagctaaaa acagttccag
     1141 cgcacgaact gcgtcattcc tgaaacttgc acaattgaca tttaatgaac cgccagagct
     1201 acttgggttg gcaattaaca gaacaaacat gaaatactcg ccgtcaacta gcgcggttgt
     1261 tagaggtgca gaagtctttc ctcgcagttt actacggacg cactccatcc cctgctgtcc
     1321 tttgtgtctg cgagaaaatg gctacgcctc ctacctttgg cactttcagg ggtacgaata
     1381 ctgccacagc cataacgtac ctttaattac cacttgtagc tgtggtaagg agtttgacta
     1441 ccgagtatct gggttaaagg gcatttgctg caaatgcaag gagcctatca ccttaaccag
     1501 cagggagaac ggtcatgagg cagcgtgtac tgtttcaaac tggcttgctg gccatgaatc
     1561 taaacctctg ccaaatcttc ctaaaagcta ccgatggggt ttagttcatt ggtggatggg
     1621 tattaaagat agcgagttcg atcacttttc gttcgttcaa tttttctcaa actggccaag
     1681 gtcattccac tcgataatcg aagatgaagt agagttcaac cttgagcatg ctgttgtcag
     1741 cacgtctgaa ttacgactaa aagatcttct tggtcgattg tttttcggtt caattcggtt
     1801 acctgagcgg aatcttcaac acaatatcat ccttggtgag cttctctgct atttagaaaa
     1861 tcgtttatgg caagacaagg gattaatcgc caacctcaaa atgaacgcgt tagaggcgac
     1921 tgtaatgtta aattgtagcc tcgatcagat tgcatcaatg gttgaacaac gcatcttgaa
     1981 gccaaatcga aaaagcaagc ccaacagccc tcttgatgtt accgattatc tatttcattt
     2041 cggcgatatt ttctgtcttt ggttagctga gttccaaagc gatgagttta accgttcgtt
     2101 ttatgtgtcg aggtggtaaa tgcaaactct gaaagaacta atcgcatcca atcctgacga
     2161 cttaacaact gaacttaaga gagcatttcg tccactcaca ccgcatattg caattgatgg
     2221 taatgaactt gacgcactga cgatattagt caatttaacc gataagactg atgatcagaa
     2281 agacctgctc gatcgagcca aatgcaagca aaaacttcga gatgaaaaat ggtgggctag
     2341 ctgcataaat tgcgttaatt acagacaaag ccataaccca aaattcccgg atatacgttc
     2401 tgaaggcgtg attcgaaccc aagccctggg tgaattaccg agtttcctac tctcgtcctc
     2461 aaaaatccca ccataccatt ggtcatatag ccatgattca aaatacgtca acaaaagcgc
     2521 attcctcacc aatgagtttt gttgggatgg tgagatctca tgtttgggtg agcttcttaa
     2581 agatgcagat cacccacttt ggaatacttt gaaaaagtta ggttgttctc aaaaaacttg
     2641 caaagcaatg gcaaaacaac tagctgatat tactctcacg actatcaatg tcacactcgc
     2701 accaaattac ctgactcaaa tatctctccc cgatagtgat acatcctaca tttcactctc
     2761 tcctgttgca tcgctatcga tgcaaagcca ctttcatcag aggctccaag atgaaaatcg
     2821 gcatagcgcg ataacgcggt ttagccgaac taccaacatg ggggtcactg cgatgacatg
     2881 tggcggcgca tttagaatgt taaagtctgg cgctaagttt tccagccccc ctcatcaccg
     2941 attaaacagt aaacgaagtt ggttgacgtc agagcatgtt cagtcattaa aacagtacca
     3001 gcgcctcaac aaaagcctca tacctgaaaa ctctcggatt gcactccgta gaaaatacaa
     3061 aatcgagctt caaaatatgg tcagatcttg gtttgcaatg caagaccata cccttgattc
     3121 gaacatactt atccaacatt tgaatcatga cctatcttac ttaggagcca caaaacgttt
     3181 tgcatacgat ccagcgatga ccaagctctt tactgagctt ttgaaacgag agttatcaaa
     3241 ttcaatcaat aatggtgagc aacacactaa tggatcgttt ttagtcctac cgaatatcag
     3301 ggtttgtggc gcaacagctt taagctcccc ggtaacggtg gggattccat cacttacagc
     3361 tttctttggc ttcgttcacg catttgaacg gaatataaat cgcaccacct catcgtttcg
     3421 tgttgaatcc tttgcgatat gcgtccatca actacatgtc gagaagcgag gtttgacagc
     3481 agagtttgtg gaaaaaggcg acgggactat atccgctccc gcgacccggg atgactggca
     3541 gtgtgatgtc gtatttagcc ttattttgaa caccaacttt gctcaacata ttgaccaaga
     3601 tacgttagtt acatcactac caaagcgatt ggctcggggt tcagcaaaaa ttgcgattga
     3661 tgactttaaa catatcaact cattctcgac attagaaaca gcgatcgaat ctctgccaat
     3721 agaagctggt aggtggttat cactttacgc acagtcaaac aataatctaa gtgatctatt
     3781 agcagccatg acagaggacc atcagctcat ggcaagctgc gtcggttacc acttgttaga
     3841 agagcccaaa gataaaccaa actccctcag aggttacaaa cacgctatcg ccgagtgcat
     3901 cattggactc attaactcaa tcacctttag ctcagagact gatcccaaca caatcttttg
     3961 gtcgctaaag aactatcaaa actacctagt ggtacagcca aggagtatca acgatgaaac
     4021 taccgacaaa tctagcctat gagcgctcta tcgacccatc agatgtctgt ttttttgtcg
     4081 tctggcccga tgatagaaaa acacctttaa cctacaattc tcgtactctg ctcgggcaaa
     4141 tggaagcggc atcattagcc tatgatgtct caggtcaacc aataaaaagt gccaccgctg
     4201 aggcgttagc tcaagggaac cctcatcaag ttgatttctg ccacgttcca tacggtgcga
     4261 gtcatattga atgcagtttc tccgtctcgt tttcttctga actacgtcaa ccatataagt
     4321 gtaactcaag caaagttaaa caaacgctag tgcaattagt cgagctctac gaaacgaaaa
     4381 tcggctggac tgagctagca acccgatatt tgatgaacat ttgcaacggt aaatggctgt
     4441 ggaaaaatac ccgtaaagcc tattgctgga acattgtact tacaccttgg ccttggaacg
     4501 gggaaaaggt tggatttgaa gatattcgta ctaactacac ctcacggcaa gactttaaaa
     4561 ataataaaaa ttggtctgct atagttgaaa tgatcaaaac cgcattttct agtactgatg
     4621 ggctggcgat atttgaagtc agggccacct tgcacttgcc aacgaatgct atggtgcggc
     4681 caagccaagt tttcacagaa aaagaaagtg gcagtaaaag taaatctaaa actcaaaaca
     4741 gtcgagtttt tcagagtaca actattgatg gtgaacgatc gccaatacta ggggccttta
     4801 aaacgggagc agctattgca accattgacg actggtatcc tgaagccact gagccactaa
     4861 gggtcggacg gtttggggtt catcgcgaag atgtcacttg ctaccgtcat ccgtctaccg
     4921 gaaaagattt tttctcgata ttacaacaag cagagcacta tattgaagtg ttgagcgcca
     4981 acaaaactcc cgctcaagaa actatcaacg acatgcactt tttaatggct aacctgatta
     5041 agggtgggat gttccagcat aaaggagact gactgtgaaa tggtattata agacaatcac
     5101 ctttctgcca gagttgtgca acaacgagtc actggctgca aagtgtctcc gcgttctgca
     5161 tggatttaac tatcagtatg agacacgaaa tatcggcgtt tcatttccgc tttggtgtga
     5221 tgcaacggtt ggaaaaaaga tttcatttgt cagcaagaac aagatagaac tcgacttact
     5281 acttaaacaa cactatttcg tccaaatgga acaacttcaa tactttcata tatccaacac
     5341 tgttctcgtc ccagaagatt gtacatacgt ttcctttaga cgctgtcaat ctatagataa
     5401 gctcacagca gcagggctgg caaggaaaat cagacgcctg gagaaacgtg ctctatctag
     5461 aggcgagcaa tttgacccat catcttttgc tcaaaaagag catactgcaa tagcgcacta
     5521 ccactcactt ggggagtcca gcaaacagac gaaccgcaac tttcgactca atatcaggat
     5581 gctctcggag caaccccgtg aggggaactc gatttttagt agctatggct tatcaaattc
     5641 agaaaactcg tttcagcctg tacccttaat ctgacctagg ctgctgccac cgctgagcaa
     5701 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct
     5761 caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag
     5821 caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg
     5881 ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg
     5941 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     6001 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     6061 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     6121 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     6181 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     6241 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     6301 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     6361 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     6421 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     6481 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     6541 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     6601 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     6661 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc
     6721 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6781 gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg
     6841 agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag
     6901 gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc
     6961 gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga
     7021 ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg
     7081 tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac
     7141 ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta
     7201 tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca
     7261 gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg
     7321 taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta
     7381 aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg
     7441 gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc
     7501 ttcgatcaaa ccacctcccc aggtggtttt ttcgaaggtt aagtcagttg gggaactgct
     7561 taaccttcga tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg
     7621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt
     7681 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     7741 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     7801 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttacc gttgtaaaac
     7861 gacggccagt gaattcctga tgaatcccct aatgattttt atcaaaatca ttaaggttac
     7921 catcacggaa aaaggttatg ctgcttttaa gacccacttt cacatttaag ttgtttttct
     7981 aatccgcata tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca
     8041 aataattcga tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt
     8101 tcttctttag cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc
     8161 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
     8221 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
     8281 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
     8341 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
     8401 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
     8461 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
     8521 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat
//
