LOCUS       sequence-340546-        6320 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 6320)
  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 6320)
  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..6320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(316..942)
                     /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     847..866
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        958..1013
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    964..982
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    994..1012
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(4518..4536)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4532..4579
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4809..5624)
                     /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(4809..5624)
                     /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     5532..5551
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5625..5716)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5734..6320,1..49))
                     /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(5879..5898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgatcgtaa aaggatctca agaagatcct ttacggattc ccgacaccat cactctagat
       61 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
      121 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
      181 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
      241 ttgcgctgat gaatccccta atgattttta tcaaaatcat taaggttacc atcacggaaa
      301 aaggttatgc tgcttttaag acccactttc acatttaagt tgtttttcta atccgcatat
      361 gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgatcaa ataattcgat
      421 agcttgtcgt aataatggcg gcatactatc agtagtaggt gtttcccttt cttctttagc
      481 gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca cagcgctgag
      541 tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta attgattttc
      601 gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg
      661 atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat cttgccagct
      721 ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa tggctaaggc
      781 gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct ctacacctag
      841 cttctgggcg agtttacggg ttgttaaacc ttcgattccg acctcattaa gcagctctaa
      901 tgcgctgtta atcactttac ttttatctaa tctagacatc attaattcct aatttttgtt
      961 gacactctat cattgataga gttattttac cactccctat cagtgataga gaaaagtgaa
     1021 ctctagaaat aattttgttt aactttaaaa ggagatatac catgtaccgt cgtaagctga
     1081 aatatagccg tgttaagaac ctgcacaaat ttgcgagcca gaagaacaaa agcacctgcc
     1141 tggtggagag cagcctggaa ttcgacgcgt gcttccactt tgagttcagc ccgccgatcg
     1201 cggcgtttga agcgcaaccg ctgggttacg agtatgaatt cgataaccgt atttgccgtt
     1261 acaccccgga ctttctgctg acccacaccg atggcaccca gaagttcatc gaggttaagc
     1321 cgcaaagcaa aattgcggac gaggattttc gtgcgcgttt catcgaaaag caggcgattg
     1381 cgaaacaaga cggtcgtgat ctgatcctgg tgaccgacaa gcagattcgt gtttacccga
     1441 ccctgaacaa cctgaaactg ctgcaccgtt atagcggctt tcagagcctg accgagctgc
     1501 aagcgagcgt gctggaactg gttaagcagt acggtagcat caaagtgggc caactgattc
     1561 gttatctgaa agttaccgcg ggtgaactgc tggcgaccgt gctgcgtctg ctgagcctgg
     1621 gccaactgtt cgcggatctg accaccaacg agatcagcat tgaaaccgcg atctggagca
     1681 acaatgttta ataacgacct gttcgacgat gagtttaacc agccgctgcc gaaggcggaa
     1741 accaaactgc cgcagaacta taccaaggat ctgcaagcgc tgccggagaa gatcaaaacc
     1801 accaccttcg cgaagctgaa atacattcaa tggctggagg cgaacatcca gggtggctgg
     1861 acccaaaaga acctggaacc gctgctgaaa ctgatgccgg acgttgaggg tgaaaagaaa
     1921 ccgagctggc gtaccgcggc gcgttggtat agcgcgtaca ccaacgcgga taagaacatt
     1981 atggcgctga tcccgagcca ccagaagaaa ggcaaccgtg aacgtgacac caccaccgat
     2041 aagttctttg agaaagcgct ggaacgttac ctggtgaagg agaaaccgag cgttgcgagc
     2101 gcgtataagt tctacaaaga cctggtgatc attgaaaacg acagcgtggt tgatagcgtt
     2161 ctgaaaccgc tgacctataa ggcgtttaaa aaccgtattg acaacctgcc gcagtatgag
     2221 gttatgatcg cgcgttacgg caagcgtctg gcggatattg cgtacaacaa ggtggaaggc
     2281 cacaaacgtc cgattcgtgt gctggagaaa gttgaaatcg accacacccc gctggatctg
     2341 attctgctgg acgatgagct gcacatcccg ctgggtcgtc cgaccctgac catgctggtt
     2401 gacgtttata gccactgcat cgtgggctac tatttcagct ttagcgagcc gagctacgat
     2461 gcggttcgtc gtgcgatgct gaacgcgatg aagccgaaaa gcgaagtggc gaaactgtac
     2521 ccggacacca ttaacgagtg gaagtgcgcg ggtaaaatcg aaaccctggt ggttgataac
     2581 ggcgcggagt tctggagcaa cagcctggaa ctggcgtgcg aggaaatcgg tattaacacc
     2641 cagtataacc cggtggcgaa gccgtggctg aaaccgttcg ttgagcgtat gtttggcacc
     2701 atcaacaccg aactgctgga cccggttccg ggcaagacct tcagcaacat cctgcaaaaa
     2761 cacgaataca acccgaagaa agacgcgatt atgcgtttca ccacctttat gcagctgttt
     2821 cacaagtggg tggttgatgt gtatcaccaa gacgcggata gccgtttcaa atacattccg
     2881 agccagctgt gggaccaagg ctttaacacc ctgccgccga ccatgctgag cgatgcggat
     2941 ctgcagcaac tggatgtggt tctgagcatc agcaaccacc gtgtgctgcg taagggtggc
     3001 attcgtctgg agaacctgag ctatgacagc accgaactgg cgaactaccg taagcagttc
     3061 agccacaaag tgagccaaga ggttctgatc aaactgaacc cggacgatat tagctacatc
     3121 tatgtgtacc tggacaagct ggaacactat attaaagttc cgtgcatcga tccgaacggt
     3181 tacacccaga acctgagcct gaaccaacac aagatcaaca ttcgtatcca ccgtgacttt
     3241 attagcggta gcatcgataa cgttggcctg gcgaaggcgc gtatgttcat tcacaacaaa
     3301 atccagaacg agtttgagga actgaagaac gcgccgaaac acagcaaggt gaaaggtggc
     3361 aaggcgctgg cgaaacacca gaacattagc agcgacagcc aaaagagcat cacccacagc
     3421 aaaccggtgg aggcgaagaa agttaccccg aaagaacaac cgaccgatag ctgggacgat
     3481 ttcatcagcg acctggatgg tttttaatta tgctgaccga caagcagaaa gaaaagctga
     3541 acgagttccg tgatgttttt attgaatacc cgatcattac caccatcttc aacgactttg
     3601 atcgtctgcg tctgggtaaa ggcctgaccg gcgagaagcc gtgcatgctg ctgaacggtg
     3661 acaccggcac cggtaaaacc gcgctgatta aacagtataa ggaacgtcac ctgccgcaat
     3721 tcatcaacgg tgttatgaac cacccggtgc tggttagccg tattccgagc aacccgaccc
     3781 tggaaagcac cctggcggag ctgctgaaag acctgggtca agtgggcagc accgagcgta
     3841 agctgcgtat taacggcacc cgtctgacca ccagcctgat caaatgcctg aagacctgcg
     3901 gcaccgaact gatcattatc gatgagtttc aggaactgat tgagcacaac caaggcaaga
     3961 aacgtcgtga aattgcgaac cgtctgaaat acatcaacga cgaggcgggt gttagcattg
     4021 tgctggttgg catgccgtgg gcggaaaaga tcgcggatga gccgcagtgg agcagccgtc
     4081 tgctgatccg tcgtcaactg ccgtatttca aactgagcga gaacccgaag cactttgtgc
     4141 agctgattat cggtctggcg aaccgtatgc cgttcgcgga aaaaccgaac ctgagcgagc
     4201 aagcgaccgt tttcaccctg tttagcctga gcaaaggctg cttccgtacc ctgaagtact
     4261 ttctggacga tgcggtgctg tatgcgctga tggacaacgc gaagaccctg accaccaaac
     4321 acctggtgaa ggcgttcgaa gttctgtttc cggatgtgcc gaacctgttt accctgccgg
     4381 ttgcggagat caccgcgagc gaggtggaac gttacagcct gtataagccg gaaagcagcc
     4441 aggacgagga cccgttcatt gcgaccaaat ttaccgatcg tatgccgatc agccaactgc
     4501 tgcgtaagta actcgagccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4561 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct
     4621 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     4681 ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg
     4741 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4801 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     4861 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     4921 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     4981 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5041 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     5101 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5161 cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca
     5221 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5281 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     5341 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     5401 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     5461 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     5521 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     5581 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     5641 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5701 tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg ccaggaggat
     5761 acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct ccgcccccct
     5821 gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     5881 agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc cgtcctgcgg
     5941 cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt gtagacagtt
     6001 cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg cgccttatcc
     6061 ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact ggcagcagcc
     6121 attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt ggcctaacag
     6181 aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag ttaccaggtt
     6241 aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg gttttttcgt
     6301 ttacagagca ggagattacg
//
