LOCUS       sequence-347981-        8478 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of CRISPR-associated transposases.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8478)
  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 8478)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..8478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        497..552
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    503..521
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    533..551
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5356..5374)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5370..5417
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5588..6379)
                     /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(6380..6471)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7548..8174)
                     /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     8079..8098
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        8190..8245
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    8196..8214
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    8226..8244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tgaactgccg agtaggtagc tgataacgtc agagtggcgt atccgatgaa tcaccacagg
       61 tgaactgccg agtaggtagc tgataacgca gaagtgcacg gcgtgtgcgg cggcgagcag
      121 tgaactgccg agtaggtagc tgataactcg atccgtgggt gtttatgttc tcagcaggag
      181 tgaactgccg agtaggtagc tgataactta cttacggcgt tgcgcattct cattgcaccg
      241 tgaactgccg agtaggtagc tgataacgca ttactaagct ggcgcgtgaa gctgtagaag
      301 tgaactgccg agtaggtagc tgataacggg gcgtaaaaag aaggggcgct gattaacgcg
      361 tgaactgccg agtaggtagc tgataacgtt taaactaggg ataacagggt aatggatccg
      421 aattcgagct cggcgcgcct gcaggtcgac aagcttgcgg ccgctctaat ctagacatca
      481 ttaattccta atttttgttg acactctatc attgatagag ttattttacc actccctatc
      541 agtgatagag aaaagtgaac tctagaaata attttgttta actttaaagg agatatacat
      601 atgtttttgc aaagacctaa accttacagc gatgaaagtt tagaaagttt ctttatccga
      661 gtggctaaca aaaatggcta cggtgatgtc catcgcttcc tagaagccac taaacgattc
      721 cttcaagaca ttgaccataa tggctatcaa acctttccga ctgatataac tcggataaac
      781 ccatactcag ctaaaaacag ttccagcgca cgaactgcgt cattcctgaa acttgcacaa
      841 ttgacattta atgaaccgcc agagctactt gggttggcaa ttaacagaac aaacatgaaa
      901 tactcgccgt caactagcgc ggttgttaga ggtgcagaag tctttcctcg cagtttacta
      961 cggacgcact ccatcccctg ctgtcctttg tgtctgcgag aaaatggcta cgcctcctac
     1021 ctttggcact ttcaggggta cgaatactgc cacagccata acgtaccttt aattaccact
     1081 tgtagctgtg gtaaggagtt tgactaccga gtatctgggt taaagggcat ttgctgcaaa
     1141 tgcaaggagc ctatcacctt aaccagcagg gagaacggtc atgaggcagc gtgtactgtt
     1201 tcaaactggc ttgctggcca tgaatctaaa cctctgccaa atcttcctaa aagctaccga
     1261 tggggtttag ttcattggtg gatgggtatt aaagatagcg agttcgatca cttttcgttc
     1321 gttcaatttt tctcaaactg gccaaggtca ttccactcga taatcgaaga tgaagtagag
     1381 ttcaaccttg agcatgctgt tgtcagcacg tctgaattac gactaaaaga tcttcttggt
     1441 cgattgtttt tcggttcaat tcggttacct gagcggaatc ttcaacacaa tatcatcctt
     1501 ggtgagcttc tctgctattt agaaaatcgt ttatggcaag acaagggatt aatcgccaac
     1561 ctcaaaatga acgcgttaga ggcgactgta atgttaaatt gtagcctcga tcagattgca
     1621 tcaatggttg aacaacgcat cttgaagcca aatcgaaaaa gcaagcccaa cagccctctt
     1681 gatgttaccg attatctatt tcatttcggc gatattttct gtctttggtt agctgagttc
     1741 caaagcgatg agtttaaccg ttcgttttat gtgtcgaggt ggtaaatgca aactctgaaa
     1801 gaactaatcg catccaatcc tgacgactta acaactgaac ttaagagagc atttcgtcca
     1861 ctcacaccgc atattgcaat tgatggtaat gaacttgacg cactgacgat attagtcaat
     1921 ttaaccgata agactgatga tcagaaagac ctgctcgatc gagccaaatg caagcaaaaa
     1981 cttcgagatg aaaaatggtg ggctagctgc ataaattgcg ttaattacag acaaagccat
     2041 aacccaaaat tcccggatat acgttctgaa ggcgtgattc gaacccaagc cctgggtgaa
     2101 ttaccgagtt tcctactctc gtcctcaaaa atcccaccat accattggtc atatagccat
     2161 gattcaaaat acgtcaacaa aagcgcattc ctcaccaatg agttttgttg ggatggtgag
     2221 atctcatgtt tgggtgagct tcttaaagat gcagatcacc cactttggaa tactttgaaa
     2281 aagttaggtt gttctcaaaa aacttgcaaa gcaatggcaa aacaactagc tgatattact
     2341 ctcacgacta tcaatgtcac actcgcacca aattacctga ctcaaatatc tctccccgat
     2401 agtgatacat cctacatttc actctctcct gttgcatcgc tatcgatgca aagccacttt
     2461 catcagaggc tccaagatga aaatcggcat agcgcgataa cgcggtttag ccgaactacc
     2521 aacatggggg tcactgcgat gacatgtggc ggcgcattta gaatgttaaa gtctggcgct
     2581 aagttttcca gcccccctca tcaccgatta aacagtaaac gaagttggtt gacgtcagag
     2641 catgttcagt cattaaaaca gtaccagcgc ctcaacaaaa gcctcatacc tgaaaactct
     2701 cggattgcac tccgtagaaa atacaaaatc gagcttcaaa atatggtcag atcttggttt
     2761 gcaatgcaag accataccct tgattcgaac atacttatcc aacatttgaa tcatgaccta
     2821 tcttacttag gagccacaaa acgttttgca tacgatccag cgatgaccaa gctctttact
     2881 gagcttttga aacgagagtt atcaaattca atcaataatg gtgagcaaca cactaatgga
     2941 tcgtttttag tcctaccgaa tatcagggtt tgtggcgcaa cagctttaag ctccccggta
     3001 acggtgggga ttccatcact tacagctttc tttggcttcg ttcacgcatt tgaacggaat
     3061 ataaatcgca ccacctcatc gtttcgtgtt gaatcctttg cgatatgcgt ccatcaacta
     3121 catgtcgaga agcgaggttt gacagcagag tttgtggaaa aaggcgacgg gactatatcc
     3181 gctcccgcga cccgggatga ctggcagtgt gatgtcgtat ttagccttat tttgaacacc
     3241 aactttgctc aacatattga ccaagatacg ttagttacat cactaccaaa gcgattggct
     3301 cggggttcag caaaaattgc gattgatgac tttaaacata tcaactcatt ctcgacatta
     3361 gaaacagcga tcgaatctct gccaatagaa gctggtaggt ggttatcact ttacgcacag
     3421 tcaaacaata atctaagtga tctattagca gccatgacag aggaccatca gctcatggca
     3481 agctgcgtcg gttaccactt gttagaagag cccaaagata aaccaaactc cctcagaggt
     3541 tacaaacacg ctatcgccga gtgcatcatt ggactcatta actcaatcac ctttagctca
     3601 gagactgatc ccaacacaat cttttggtcg ctaaagaact atcaaaacta cctagtggta
     3661 cagccaagga gtatcaacga tgaaactacc gacaaatcta gcctatgagc gctctatcga
     3721 cccatcagat gtctgttttt ttgtcgtctg gcccgatgat agaaaaacac ctttaaccta
     3781 caattctcgt actctgctcg ggcaaatgga agcggcatca ttagcctatg atgtctcagg
     3841 tcaaccaata aaaagtgcca ccgctgaggc gttagctcaa gggaaccctc atcaagttga
     3901 tttctgccac gttccatacg gtgcgagtca tattgaatgc agtttctccg tctcgttttc
     3961 ttctgaacta cgtcaaccat ataagtgtaa ctcaagcaaa gttaaacaaa cgctagtgca
     4021 attagtcgag ctctacgaaa cgaaaatcgg ctggactgag ctagcaaccc gatatttgat
     4081 gaacatttgc aacggtaaat ggctgtggaa aaatacccgt aaagcctatt gctggaacat
     4141 tgtacttaca ccttggcctt ggaacgggga aaaggttgga tttgaagata ttcgtactaa
     4201 ctacacctca cggcaagact ttaaaaataa taaaaattgg tctgctatag ttgaaatgat
     4261 caaaaccgca ttttctagta ctgatgggct ggcgatattt gaagtcaggg ccaccttgca
     4321 cttgccaacg aatgctatgg tgcggccaag ccaagttttc acagaaaaag aaagtggcag
     4381 taaaagtaaa tctaaaactc aaaacagtcg agtttttcag agtacaacta ttgatggtga
     4441 acgatcgcca atactagggg cctttaaaac gggagcagct attgcaacca ttgacgactg
     4501 gtatcctgaa gccactgagc cactaagggt cggacggttt ggggttcatc gcgaagatgt
     4561 cacttgctac cgtcatccgt ctaccggaaa agattttttc tcgatattac aacaagcaga
     4621 gcactatatt gaagtgttga gcgccaacaa aactcccgct caagaaacta tcaacgacat
     4681 gcacttttta atggctaacc tgattaaggg tgggatgttc cagcataaag gagactgact
     4741 gtgaaatggt attataagac aatcaccttt ctgccagagt tgtgcaacaa cgagtcactg
     4801 gctgcaaagt gtctccgcgt tctgcatgga tttaactatc agtatgagac acgaaatatc
     4861 ggcgtttcat ttccgctttg gtgtgatgca acggttggaa aaaagatttc atttgtcagc
     4921 aagaacaaga tagaactcga cttactactt aaacaacact atttcgtcca aatggaacaa
     4981 cttcaatact ttcatatatc caacactgtt ctcgtcccag aagattgtac atacgtttcc
     5041 tttagacgct gtcaatctat agataagctc acagcagcag ggctggcaag gaaaatcaga
     5101 cgcctggaga aacgtgctct atctagaggc gagcaatttg acccatcatc ttttgctcaa
     5161 aaagagcata ctgcaatagc gcactaccac tcacttgggg agtccagcaa acagacgaac
     5221 cgcaactttc gactcaatat caggatgctc tcggagcaac cccgtgaggg gaactcgatt
     5281 tttagtagct atggcttatc aaattcagaa aactcgtttc agcctgtacc cttaatctga
     5341 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     5401 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     5461 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     5521 ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc ccctcggctt gaacgaattg
     5581 ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt
     5641 ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag
     5701 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     5761 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     5821 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     5881 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     5941 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     6001 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     6061 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     6121 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     6181 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     6241 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     6301 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     6361 cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa atagctagct
     6481 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
     6541 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
     6601 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
     6661 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
     6721 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
     6781 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
     6841 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
     6901 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
     6961 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
     7021 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
     7081 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
     7141 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
     7201 aaggttaagt cagttgggga actgcttaac cttcgatcaa accacctccc caggtggttt
     7261 tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7321 tcttttctac tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     7381 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
     7441 gaaggagctg actgggttga aggctctcaa gggcatcctg atgaatcccc taatgatttt
     7501 tatcaaaatc attaaggtta ccatcacgga aaaaggttat gctgctttta agacccactt
     7561 tcacatttaa gttgtttttc taatccgcat atgatcaatt caaggccgaa taagaaggct
     7621 ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg cggcatacta
     7681 tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc ttccaatacg
     7741 caacctaaag taaaatgccc cacagcgctg agtgcatata atgcattctc tagtgaaaaa
     7801 ccttgttggc ataaaaaggc taattgattt tcgagagttt catactgttt ttctgtaggc
     7861 cgtgtaccta aatgtacttt tgctccatcg cgatgactta gtaaagcaca tctaaaactt
     7921 ttagcgttat tacgtaaaaa atcttgccag ctttcccctt ctaaagggca aaagtgagta
     7981 tggtgcctat ctaacatctc aatggctaag gcgtcgagca aagcccgctt attttttaca
     8041 tgccaataca atgtaggctg ctctacacct agcttctggg cgagtttacg ggttgttaaa
     8101 ccttcgattc cgacctcatt aagcagctct aatgcgctgt taatcacttt acttttatct
     8161 aatctagaca tcattaattc ctaatttttg ttgacactct atcattgata gagttatttt
     8221 accactccct atcagtgata gagaaaagtg aactctagaa ataattttgt ttaactttaa
     8281 aaggagatat accatgggtg aactgccgag taggtagctg ataactggat gctttgcgtg
     8341 caaaggaacc tgaactcgtg aactgccgag taggtagctg ataacgcctg gcttggcgat
     8401 gctcgcctga tcgacaagtg aactgccgag taggtagctg ataacgagaa tattgtggca
     8461 aaagcgcata aggtgctg
//
