LOCUS       sequence-347011-        6342 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 6342)
  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 6342)
  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: pTns(Ptr)
FEATURES             Location/Qualifiers
     source          1..6342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3308..3326)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3322..3369
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3599..4414)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3599..4414)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4322..4341
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4415..4506)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4524..5159)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(4669..4688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5419..6045)
                     /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     5950..5969
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        6061..6116
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    6067..6085
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    6097..6115
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 agcgcaaccg ctgggttacg agtatgaatt cgataaccgt atttgccgtt acaccccgga
       61 ctttctgctg acccacaccg atggcaccca gaagttcatc gaggttaagc cgcaaagcaa
      121 aattgcggac gaggattttc gtgcgcgttt catcgaaaag caggcgattg cgaaacaaga
      181 cggtcgtgat ctgatcctgg tgaccgacaa gcagattcgt gtttacccga ccctgaacaa
      241 cctgaaactg ctgcaccgtt atagcggctt tcagagcctg accgagctgc aagcgagcgt
      301 gctggaactg gttaagcagt acggtagcat caaagtgggc caactgattc gttatctgaa
      361 agttaccgcg ggtgaactgc tggcgaccgt gctgcgtctg ctgagcctgg gccaactgtt
      421 cgcggatctg accaccaacg agatcagcat tgaaaccgcg atctggagca acaatgttta
      481 ataacgacct gttcgacgat gagtttaacc agccgctgcc gaaggcggaa accaaactgc
      541 cgcagaacta taccaaggat ctgcaagcgc tgccggagaa gatcaaaacc accaccttcg
      601 cgaagctgaa atacattcaa tggctggagg cgaacatcca gggtggctgg acccaaaaga
      661 acctggaacc gctgctgaaa ctgatgccgg acgttgaggg tgaaaagaaa ccgagctggc
      721 gtaccgcggc gcgttggtat agcgcgtaca ccaacgcgga taagaacatt atggcgctga
      781 tcccgagcca ccagaagaaa ggcaaccgtg aacgtgacac caccaccgat aagttctttg
      841 agaaagcgct ggaacgttac ctggtgaagg agaaaccgag cgttgcgagc gcgtataagt
      901 tctacaaaga cctggtgatc attgaaaacg acagcgtggt tgatagcgtt ctgaaaccgc
      961 tgacctataa ggcgtttaaa aaccgtattg acaacctgcc gcagtatgag gttatgatcg
     1021 cgcgttacgg caagcgtctg gcggatattg cgtacaacaa ggtggaaggc cacaaacgtc
     1081 cgattcgtgt gctggagaaa gttgaaatcg accacacccc gctggatctg attctgctgg
     1141 acgatgagct gcacatcccg ctgggtcgtc cgaccctgac catgctggtt gacgtttata
     1201 gccactgcat cgtgggctac tatttcagct ttagcgagcc gagctacgat gcggttcgtc
     1261 gtgcgatgct gaacgcgatg aagccgaaaa gcgaagtggc gaaactgtac ccggacacca
     1321 ttaacgagtg gaagtgcgcg ggtaaaatcg aaaccctggt ggttgataac ggcgcggagt
     1381 tctggagcaa cagcctggaa ctggcgtgcg aggaaatcgg tattaacacc cagtataacc
     1441 cggtggcgaa gccgtggctg aaaccgttcg ttgagcgtat gtttggcacc atcaacaccg
     1501 aactgctgga cccggttccg ggcaagacct tcagcaacat cctgcaaaaa cacgaataca
     1561 acccgaagaa agacgcgatt atgcgtttca ccacctttat gcagctgttt cacaagtggg
     1621 tggttgatgt gtatcaccaa gacgcggata gccgtttcaa atacattccg agccagctgt
     1681 gggaccaagg ctttaacacc ctgccgccga ccatgctgag cgatgcggat ctgcagcaac
     1741 tggatgtggt tctgagcatc agcaaccacc gtgtgctgcg taagggtggc attcgtctgg
     1801 agaacctgag ctatgacagc accgaactgg cgaactaccg taagcagttc agccacaaag
     1861 tgagccaaga ggttctgatc aaactgaacc cggacgatat tagctacatc tatgtgtacc
     1921 tggacaagct ggaacactat attaaagttc cgtgcatcga tccgaacggt tacacccaga
     1981 acctgagcct gaaccaacac aagatcaaca ttcgtatcca ccgtgacttt attagcggta
     2041 gcatcgataa cgttggcctg gcgaaggcgc gtatgttcat tcacaacaaa atccagaacg
     2101 agtttgagga actgaagaac gcgccgaaac acagcaaggt gaaaggtggc aaggcgctgg
     2161 cgaaacacca gaacattagc agcgacagcc aaaagagcat cacccacagc aaaccggtgg
     2221 aggcgaagaa agttaccccg aaagaacaac cgaccgatag ctgggacgat ttcatcagcg
     2281 acctggatgg tttttaatta tgctgaccga caagcagaaa gaaaagctga acgagttccg
     2341 tgatgttttt attgaatacc cgatcattac caccatcttc aacgactttg atcgtctgcg
     2401 tctgggtaaa ggcctgaccg gcgagaagcc gtgcatgctg ctgaacggtg acaccggcac
     2461 cggtaaaacc gcgctgatta aacagtataa ggaacgtcac ctgccgcaat tcatcaacgg
     2521 tgttatgaac cacccggtgc tggttagccg tattccgagc aacccgaccc tggaaagcac
     2581 cctggcggag ctgctgaaag acctgggtca agtgggcagc accgagcgta agctgcgtat
     2641 taacggcacc cgtctgacca ccagcctgat caaatgcctg aagacctgcg gcaccgaact
     2701 gatcattatc gatgagtttc aggaactgat tgagcacaac caaggcaaga aacgtcgtga
     2761 aattgcgaac cgtctgaaat acatcaacga cgaggcgggt gttagcattg tgctggttgg
     2821 catgccgtgg gcggaaaaga tcgcggatga gccgcagtgg agcagccgtc tgctgatccg
     2881 tcgtcaactg ccgtatttca aactgagcga gaacccgaag cactttgtgc agctgattat
     2941 cggtctggcg aaccgtatgc cgttcgcgga aaaaccgaac ctgagcgagc aagcgaccgt
     3001 tttcaccctg tttagcctga gcaaaggctg cttccgtacc ctgaagtact ttctggacga
     3061 tgcggtgctg tatgcgctga tggacaacgc gaagaccctg accaccaaac acctggtgaa
     3121 ggcgttcgaa gttctgtttc cggatgtgcc gaacctgttt accctgccgg ttgcggagat
     3181 caccgcgagc gaggtggaac gttacagcct gtataagccg gaaagcagcc aggacgagga
     3241 cccgttcatt gcgaccaaat ttaccgatcg tatgccgatc agccaactgc tgcgtaagta
     3301 actcgagccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3361 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     3421 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     3481 acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg tgcgcggaac
     3541 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
     3601 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     3661 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     3721 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     3781 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     3841 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     3901 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     3961 cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat
     4021 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     4081 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     4141 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     4201 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     4261 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     4321 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     4381 tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa
     4441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4501 aacaaatagg catgctagcg cagaaacgtc ctagaagatg ccaggaggat acttagcaga
     4561 gagacaataa ggccggagcg aagccgtttt tccataggct ccgcccccct gacgaacatc
     4621 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     4681 cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc cgtcctgcgg cgtccgtgtt
     4741 gtggtggagg ctttacccaa atcaccacgt cccgttccgt gtagacagtt cgctccaagc
     4801 tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg cgccttatcc ggtaactatc
     4861 atcttgagtc caacccggaa agacacgaca aaacgccact ggcagcagcc attggtaact
     4921 gagaattagt ggatttagat atcgagagtc ttgaagtggt ggcctaacag aggctacact
     4981 gaaaggacag tatttggtat ctgcgctcca ctaaagccag ttaccaggtt aagcagttcc
     5041 ccaactgact taaccttcga tcaaaccgcc tccccaggcg gttttttcgt ttacagagca
     5101 ggagattacg acgatcgtaa aaggatctca agaagatcct ttacggattc ccgacaccat
     5161 cactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     5221 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     5281 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5341 attaattgcg ttgcgcctgc ctaatgattt ttatcaaaat cattaaggtt accatcacgg
     5401 aaaaaggtta tgctgctttt aagacccact ttcacattta agttgttttt ctaatccgca
     5461 tatgatcaat tcaaggccga ataagaaggc tggctctgca ccttggtgat caaataattc
     5521 gatagcttgt cgtaataatg gcggcatact atcagtagta ggtgtttccc tttcttcttt
     5581 agcgacttga tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct
     5641 gagtgcatat aatgcattct ctagtgaaaa accttgttgg cataaaaagg ctaattgatt
     5701 ttcgagagtt tcatactgtt tttctgtagg ccgtgtacct aaatgtactt ttgctccatc
     5761 gcgatgactt agtaaagcac atctaaaact tttagcgtta ttacgtaaaa aatcttgcca
     5821 gctttcccct tctaaagggc aaaagtgagt atggtgccta tctaacatct caatggctaa
     5881 ggcgtcgagc aaagcccgct tattttttac atgccaatac aatgtaggct gctctacacc
     5941 tagcttctgg gcgagtttac gggttgttaa accttcgatt ccgacctcat taagcagctc
     6001 taatgcgctg ttaatcactt tacttttatc taatctagac atcattaatt cctaattttt
     6061 gttgacactc tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt
     6121 gaactctaga aataattttg tttaacttta acctgtagaa ataattttgt ttaactttaa
     6181 aaggagatat accatgtacc gtcgtaagct gaaatatagc cgtgttaaga acctgcacaa
     6241 atttgcgagc cagaagaaca aaagcacctg cctggtggag agcagcctgg aattcgacgc
     6301 gtgcttccac tttgagttca gcccgccgat cgcggcgttt ga
//
