LOCUS       sequence-338902-        6403 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 6403)
  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 6403)
  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: pTet-tns
FEATURES             Location/Qualifiers
     source          1..6403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     318..337
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        429..484
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    435..453
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    465..483
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2641..2652
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4079..4097)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4093..4140
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4370..5185)
                     /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(4370..5185)
                     /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     5093..5112
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5186..5277)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5295..5930)
                     /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(5440..5459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6190..6403,1..413))
                     /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"
ORIGIN
        1 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
       61 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
      121 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
      181 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
      241 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
      301 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
      361 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat cattaattcc
      421 taatttttgt tgacactcta tcattgatag agttatttta ccactcccta tcagtgatag
      481 agaaaagtga actctagaaa taattttgtt taactttaac ctgtagaaat aattttgttt
      541 aactttaata aggagatata ccatggcgac aagtttacct acgccctcag caattacgac
      601 ttcggcgtta gagtatgcat tccatactcc cgctcgcaat ctaacgaaat ctcgcggaaa
      661 aaacattcat cgttatgtca gtgtaaagat gagtaagagg attacggtag aatctactct
      721 agagtgtgat gcctgctatc actttgattt tgagccaagt attgttcgct tttgcgctca
      781 accgattcga tttttatatt atctcaatgg tcagtctcac tcctatgttc ctgactttct
      841 agttcaattt gataccaacg agtttgttct atatgaagta aagtcagctt atgctaagaa
      901 caaacctgat tttgatgttg aatgggaggc gaaagtaaaa gcagcaactg aactagggct
      961 agaattggag cttgttgaag agagtgatat tagggatacg gttgtattaa ataatcttaa
     1021 gcgcatgcat cgttatgctt cgaaagatga gctgaataac gtacataact ctctcttaaa
     1081 aataataaag tacaatggcg cccaatctgc aagatgcttg ggagaacagt tgggtttaaa
     1141 aggccgaact gttttaccaa ttttgtgcga tttgctgtca aggtgtttac tcgatacacg
     1201 tttggataag cctctatctc ttgaatctcg atttgagttg gccagttatg gctaagaaag
     1261 ggttctcaag tttccataga aaagcagtct cttcccaaga tacgcttgaa tcgatagagc
     1321 ttgtctctag cgctaattgt ctagaaagtg ttacgtatca agatatatca gcatttcccg
     1381 aaacaattgc ggtagagatt aatttccgat taagcattct tcgcttttta gcgcgaaagt
     1441 gcgaaaccat tgtggccaaa tcaattgaac cacatcgtgt agagctacag caaaactata
     1501 gtagaaaaat acccagtgca ataacgatat atcgatggtg gcttgctttt cgaaaatcag
     1561 actacaaccc cattagctta gcacctaata tcaaggatag aggtaataga gagacaaaag
     1621 tgtcaacagt tgttgattct attatggaac aggcagttga aagagttata tctggacgaa
     1681 aagtcaatgt tagctctgca tataaacgtg ttcgacgaaa agttcgtcaa tacaatctaa
     1741 ctcatggaac gaaatacacg tatcctaagt acgaatctgt aagaaagcga gtaaaaaaga
     1801 aaaccccatt tgagttatta gccgcaggga aaggggagag agtagctaag agagagtttc
     1861 gccgaatggg aaaaaagatc ctcacgtcta gcgtgctaga gagggttgaa atagatcaca
     1921 ctgtcgttga cctttttgca gtacacgaag agtatcgaat cccattgggc cgaccttggc
     1981 ttactcaatt ggttgattgt tacagtaaag ctgtaatcgg tttttattta ggtttcgagc
     2041 ctcctagcta tgtgtcggtt tcccttgcac ttaagaatgc aatacaacgc aaagatgact
     2101 taatctcctc gtatgaatcg atcgagaatg aatggctatg ttatggcatc ccagacctac
     2161 tcgtaactga taatggtaaa gagtttttgt cgaaagcatt tgatcaagca tgtgaatcac
     2221 tattgatcaa tgtgcatcaa aataaagttg agacgcccga caacaaacct catgttgaac
     2281 gtaactacgg gactattaat acttctctgt tagacgattt acctgggaaa tccttcagcc
     2341 agtaccttca aagagaaggg tacgactctg tgggagaagc tacccttaca ctcaatgaga
     2401 ttagagaaat ttacttaatt tggttggtgg atatttatca taaaaaaccc aatcagagag
     2461 gcactaattg tcctaatgtt gcgtggaaaa agggttgtca agaatgggaa ccagaggagt
     2521 tctctggttc taaagacgaa ttagacttta aatttgctat tgttgattac aaacaactta
     2581 ctaaagtagg gataactgtc tacaaagaac tgagttatag caatgaccgt ttagctgaat
     2641 atagagggaa gaaaggaaac cataaagttc agttcaagta taaccctgag tgtatggcag
     2701 ttatttgggt gttggatgag gatatgaatg agtactttac agttaatgcg attgactacg
     2761 aatatgcaag tagagtatca ctttggcaac ataaatataa catgaaatat caagcagaac
     2821 taaattcagc agaatatgat gaggacaagg aaattgatgc agaaataaaa attgaagaaa
     2881 tcgcagatcg ttcaattgtt aagactaaca aaatcagagc tcggaggcgt ggcgctaggc
     2941 atcaagagaa tagcgcaagg gctaagtcaa tcagtaatgc gaacccggcc tcgatacaaa
     3001 aacatgaaga tgaaatcgtt agtgcagata atgacgattg ggatattgat tatgtctgag
     3061 aaatcgtcaa atgagtgaaa cgcgtgaggc tcgtatatca agagctaaaa gggcatttgt
     3121 atccacaccg agcgttagga aaatcttgag ttacatggat agatgtagag atctatcaga
     3181 cctagagtct gagcctacat gcatgatggt ctatggtgct tcgggtgtag gtaaaacgac
     3241 cgtcatcaag aaatacttaa atcaaaacag aagagagtcc gaagccgggg gcgatataat
     3301 accggttttg catattgagt tgccagacaa tgcgaagcca gtagatgcag caagggaatt
     3361 gctggttgaa atgggtgacc cgctagcact ttatgaaact gacttagcta gattgacgaa
     3421 aagactgact gaattaatcc ctgcggtcgg cgtgaagctg attattatcg atgagttcca
     3481 acatttggtg gaagaaaggt caaatcgggt tcttacccaa gtaggtaatt ggctaaaaat
     3541 gatacttaac aaaacgaaat gtccaattgt tatatttggt atgccatact caaaagttgt
     3601 actgcaagca aactcgcaac ttcacgggcg attttccatt caggttgaac tgcgcccctt
     3661 tagctaccag ggaggtagag gtgtatttaa aacttttttg gaataccttg ataaagccct
     3721 accttttgaa aaacaggctg gcttagccaa cgaaagtttg cagaaaaaat tgtatgcatt
     3781 ctctcaggga aacatgcgtt cgttgagaaa ccttatttat caagcatcta tcgaagcaat
     3841 tgataatcag catgagacga taaccgaaga agatttcgtt tttgcatcga agttgacatc
     3901 gggcgataaa cccaactcat ggaaaaatcc ttttgaggag ggtgttgagg taacagaaga
     3961 tatgttacga ccgccaccaa aagatattgg ttgggaggac tatttgagac attcaacccc
     4021 gagagtgagt aaaccaggta gaaataaaaa ctttttcgaa taacctaggc tgctgccacc
     4081 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     4141 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     4201 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgacaagc
     4261 tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4321 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     4381 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     4441 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     4501 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     4561 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     4621 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     4681 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     4741 gacgaaatac gcggtcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     4801 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     4861 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     4921 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     4981 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     5041 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     5101 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     5161 acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc
     5221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5281 gcatgctagc gcagaaacgt cctagaagat gccaggagga tacttagcag agagacaata
     5341 aggccggagc gaagccgttt ttccataggc tccgcccccc tgacgaacat cacgaaatct
     5401 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5461 ctgatggctc cctcttgcgc tctcctgttc ccgtcctgcg gcgtccgtgt tgtggtggag
     5521 gctttaccca aatcaccacg tcccgttccg tgtagacagt tcgctccaag ctgggctgtg
     5581 tgcaagaacc ccccgttcag cccgactgct gcgccttatc cggtaactat catcttgagt
     5641 ccaacccgga aagacacgac aaaacgccac tggcagcagc cattggtaac tgagaattag
     5701 tggatttaga tatcgagagt cttgaagtgg tggcctaaca gaggctacac tgaaaggaca
     5761 gtatttggta tctgcgctcc actaaagcca gttaccaggt taagcagttc cccaactgac
     5821 ttaaccttcg atcaaaccgc ctccccaggc ggttttttcg tttacagagc aggagattac
     5881 gacgatcgta aaaggatctc aagaagatcc tttacggatt cccgacacca tcactctaga
     5941 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     6001 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     6061 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     6121 gttgcgcctg cctaatgatt tttatcaaaa tcattaaggt taccatcacg gaaaaaggtt
     6181 atgctgcttt taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
     6241 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
     6301 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
     6361 atgctcttga tcttccaata cgcaacctaa agtaaaatgc ccc
//
