pTetQCas-BsaI

LOCUS       pTetQCas-BsaI                7955 bp ds-DNA     circular SYN 03-AUG-2021
DEFINITION  Expression of CRISPR-associated transposases.
ACCESSION   .
VERSION     .
KEYWORDS    pTetQCas-BsaI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7955)
  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 7955)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 3, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7955
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        281..336
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    287..305
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    317..335
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5140..5158)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5154..5201
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5372..6163)
                     /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(6164..6255)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7332..7955)
                     /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     7863..7882
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 cattaattcc taatttttgt tgacactcta tcattgatag agttatttta ccactcccta
       61 tcagtgatag agaaaagtga actctagaaa taattttgtt taactttaaa aggagatata
      121 ccatgggtga actgccgagt aggtagctga taacgagacc tctggtctcg tgaactgccg
      181 agtaggtagc tgataacgga tccgaattcg agctcggcgc gcctgcaggt cgacaagctt
      241 gcggccgctc taatctagac atcattaatt cctaattttt gttgacactc tatcattgat
      301 agagttattt taccactccc tatcagtgat agagaaaagt gaactctaga aataattttg
      361 tttaacttta aaggagatat acatatgttt ttgcaaagac ctaaacctta cagcgatgaa
      421 agtttagaaa gtttctttat ccgagtggct aacaaaaatg gctacggtga tgtccatcgc
      481 ttcctagaag ccactaaacg attccttcaa gacattgacc ataatggcta tcaaaccttt
      541 ccgactgata taactcggat aaacccatac tcagctaaaa acagttccag cgcacgaact
      601 gcgtcattcc tgaaacttgc acaattgaca tttaatgaac cgccagagct acttgggttg
      661 gcaattaaca gaacaaacat gaaatactcg ccgtcaacta gcgcggttgt tagaggtgca
      721 gaagtctttc ctcgcagttt actacggacg cactccatcc cctgctgtcc tttgtgtctg
      781 cgagaaaatg gctacgcctc ctacctttgg cactttcagg ggtacgaata ctgccacagc
      841 cataacgtac ctttaattac cacttgtagc tgtggtaagg agtttgacta ccgagtatct
      901 gggttaaagg gcatttgctg caaatgcaag gagcctatca ccttaaccag cagggagaac
      961 ggtcatgagg cagcgtgtac tgtttcaaac tggcttgctg gccatgaatc taaacctctg
     1021 ccaaatcttc ctaaaagcta ccgatggggt ttagttcatt ggtggatggg tattaaagat
     1081 agcgagttcg atcacttttc gttcgttcaa tttttctcaa actggccaag gtcattccac
     1141 tcgataatcg aagatgaagt agagttcaac cttgagcatg ctgttgtcag cacgtctgaa
     1201 ttacgactaa aagatcttct tggtcgattg tttttcggtt caattcggtt acctgagcgg
     1261 aatcttcaac acaatatcat ccttggtgag cttctctgct atttagaaaa tcgtttatgg
     1321 caagacaagg gattaatcgc caacctcaaa atgaacgcgt tagaggcgac tgtaatgtta
     1381 aattgtagcc tcgatcagat tgcatcaatg gttgaacaac gcatcttgaa gccaaatcga
     1441 aaaagcaagc ccaacagccc tcttgatgtt accgattatc tatttcattt cggcgatatt
     1501 ttctgtcttt ggttagctga gttccaaagc gatgagttta accgttcgtt ttatgtgtcg
     1561 aggtggtaaa tgcaaactct gaaagaacta atcgcatcca atcctgacga cttaacaact
     1621 gaacttaaga gagcatttcg tccactcaca ccgcatattg caattgatgg taatgaactt
     1681 gacgcactga cgatattagt caatttaacc gataagactg atgatcagaa agacctgctc
     1741 gatcgagcca aatgcaagca aaaacttcga gatgaaaaat ggtgggctag ctgcataaat
     1801 tgcgttaatt acagacaaag ccataaccca aaattcccgg atatacgttc tgaaggcgtg
     1861 attcgaaccc aagccctggg tgaattaccg agtttcctac tctcgtcctc aaaaatccca
     1921 ccataccatt ggtcatatag ccatgattca aaatacgtca acaaaagcgc attcctcacc
     1981 aatgagtttt gttgggatgg tgagatctca tgtttgggtg agcttcttaa agatgcagat
     2041 cacccacttt ggaatacttt gaaaaagtta ggttgttctc aaaaaacttg caaagcaatg
     2101 gcaaaacaac tagctgatat tactctcacg actatcaatg tcacactcgc accaaattac
     2161 ctgactcaaa tatctctccc cgatagtgat acatcctaca tttcactctc tcctgttgca
     2221 tcgctatcga tgcaaagcca ctttcatcag aggctccaag atgaaaatcg gcatagcgcg
     2281 ataacgcggt ttagccgaac taccaacatg ggggtcactg cgatgacatg tggcggcgca
     2341 tttagaatgt taaagtctgg cgctaagttt tccagccccc ctcatcaccg attaaacagt
     2401 aaacgaagtt ggttgacgtc agagcatgtt cagtcattaa aacagtacca gcgcctcaac
     2461 aaaagcctca tacctgaaaa ctctcggatt gcactccgta gaaaatacaa aatcgagctt
     2521 caaaatatgg tcagatcttg gtttgcaatg caagaccata cccttgattc gaacatactt
     2581 atccaacatt tgaatcatga cctatcttac ttaggagcca caaaacgttt tgcatacgat
     2641 ccagcgatga ccaagctctt tactgagctt ttgaaacgag agttatcaaa ttcaatcaat
     2701 aatggtgagc aacacactaa tggatcgttt ttagtcctac cgaatatcag ggtttgtggc
     2761 gcaacagctt taagctcccc ggtaacggtg gggattccat cacttacagc tttctttggc
     2821 ttcgttcacg catttgaacg gaatataaat cgcaccacct catcgtttcg tgttgaatcc
     2881 tttgcgatat gcgtccatca actacatgtc gagaagcgag gtttgacagc agagtttgtg
     2941 gaaaaaggcg acgggactat atccgctccc gcgacccggg atgactggca gtgtgatgtc
     3001 gtatttagcc ttattttgaa caccaacttt gctcaacata ttgaccaaga tacgttagtt
     3061 acatcactac caaagcgatt ggctcggggt tcagcaaaaa ttgcgattga tgactttaaa
     3121 catatcaact cattctcgac attagaaaca gcgatcgaat ctctgccaat agaagctggt
     3181 aggtggttat cactttacgc acagtcaaac aataatctaa gtgatctatt agcagccatg
     3241 acagaggacc atcagctcat ggcaagctgc gtcggttacc acttgttaga agagcccaaa
     3301 gataaaccaa actccctcag aggttacaaa cacgctatcg ccgagtgcat cattggactc
     3361 attaactcaa tcacctttag ctcagagact gatcccaaca caatcttttg gtcgctaaag
     3421 aactatcaaa actacctagt ggtacagcca aggagtatca acgatgaaac taccgacaaa
     3481 tctagcctat gagcgctcta tcgacccatc agatgtctgt ttttttgtcg tctggcccga
     3541 tgatagaaaa acacctttaa cctacaattc tcgtactctg ctcgggcaaa tggaagcggc
     3601 atcattagcc tatgatgtct caggtcaacc aataaaaagt gccaccgctg aggcgttagc
     3661 tcaagggaac cctcatcaag ttgatttctg ccacgttcca tacggtgcga gtcatattga
     3721 atgcagtttc tccgtctcgt tttcttctga actacgtcaa ccatataagt gtaactcaag
     3781 caaagttaaa caaacgctag tgcaattagt cgagctctac gaaacgaaaa tcggctggac
     3841 tgagctagca acccgatatt tgatgaacat ttgcaacggt aaatggctgt ggaaaaatac
     3901 ccgtaaagcc tattgctgga acattgtact tacaccttgg ccttggaacg gggaaaaggt
     3961 tggatttgaa gatattcgta ctaactacac ctcacggcaa gactttaaaa ataataaaaa
     4021 ttggtctgct atagttgaaa tgatcaaaac cgcattttct agtactgatg ggctggcgat
     4081 atttgaagtc agggccacct tgcacttgcc aacgaatgct atggtgcggc caagccaagt
     4141 tttcacagaa aaagaaagtg gcagtaaaag taaatctaaa actcaaaaca gtcgagtttt
     4201 tcagagtaca actattgatg gtgaacgatc gccaatacta ggggccttta aaacgggagc
     4261 agctattgca accattgacg actggtatcc tgaagccact gagccactaa gggtcggacg
     4321 gtttggggtt catcgcgaag atgtcacttg ctaccgtcat ccgtctaccg gaaaagattt
     4381 tttctcgata ttacaacaag cagagcacta tattgaagtg ttgagcgcca acaaaactcc
     4441 cgctcaagaa actatcaacg acatgcactt tttaatggct aacctgatta agggtgggat
     4501 gttccagcat aaaggagact gactgtgaaa tggtattata agacaatcac ctttctgcca
     4561 gagttgtgca acaacgagtc actggctgca aagtgtctcc gcgttctgca tggatttaac
     4621 tatcagtatg agacacgaaa tatcggcgtt tcatttccgc tttggtgtga tgcaacggtt
     4681 ggaaaaaaga tttcatttgt cagcaagaac aagatagaac tcgacttact acttaaacaa
     4741 cactatttcg tccaaatgga acaacttcaa tactttcata tatccaacac tgttctcgtc
     4801 ccagaagatt gtacatacgt ttcctttaga cgctgtcaat ctatagataa gctcacagca
     4861 gcagggctgg caaggaaaat cagacgcctg gagaaacgtg ctctatctag aggcgagcaa
     4921 tttgacccat catcttttgc tcaaaaagag catactgcaa tagcgcacta ccactcactt
     4981 ggggagtcca gcaaacagac gaaccgcaac tttcgactca atatcaggat gctctcggag
     5041 caaccccgtg aggggaactc gatttttagt agctatggct tatcaaattc agaaaactcg
     5101 tttcagcctg tacccttaat ctgacctagg ctgctgccac cgctgagcaa taactagcat
     5161 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
     5221 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     5281 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg
     5341 cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg gtgatctcgc
     5401 ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt
     5461 cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
     5521 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
     5581 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
     5641 agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat
     5701 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
     5761 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
     5821 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
     5881 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
     5941 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
     6001 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
     6061 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
     6121 ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat
     6181 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6241 agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg agactgcggc
     6301 gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag gggactaaca
     6361 tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc gccctcctgc
     6421 cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga ggctcaacca
     6481 tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg tttccggcgg
     6541 gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac ctgttccgcc
     6601 tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta tctcggctcg
     6661 gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca gcccgactgc
     6721 tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg taaaacgcca
     6781 ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta aacacgcggt
     6841 tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg gttgctgtgc
     6901 tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc ttcgatcaaa
     6961 ccacctcccc aggtggtttt ttcgaaggtt aagtcagttg gggaactgct taaccttcga
     7021 tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa
     7081 aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat
     7141 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     7201 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     7261 cctgatgaat cccctaatga tttttatcaa aatcattaag gttaccatca cggaaaaagg
     7321 ttatgctgct tttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc
     7381 aattcaaggc cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct
     7441 tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact
     7501 tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca
     7561 tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga
     7621 gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga
     7681 cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc
     7741 ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg
     7801 agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc
     7861 tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg
     7921 ctgttaatca ctttactttt atctaatcta gacat
//