LOCUS       sequence-308952-        4504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the aTc-inducible expression of the v5.9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4504)
  AUTHORS   Milligan JN, Shroff R, Garry DJ, Ellington AD
  TITLE     Evolution of a Thermophilic Strand-Displacing Polymerase Using 
            High-Temperature Isothermal Compartmentalized Self-Replication.
  JOURNAL   Biochemistry. 2018 Aug 7;57(31):4607-4619. doi: 
            10.1021/acs.biochem.8b00200. Epub 2018 Apr 18.
  PUBMED    29629759
REFERENCE   2  (bases 1 to 4504)
  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: 
            "10.1021/acs.biochem.8b00200"; journalName: "Biochemistry"; date: 
            "2018-08-7- 7"; volume: "57"; issue: "31"; pages: "4607-4619"
COMMENT     Sequence Label: pAtetO_6xHis-v5.9
FEATURES             Location/Qualifiers
     source          1..4504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..56
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7..25
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             118..135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             136..147
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(2013..2601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2093..2112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2754..3380)
                     /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     3285..3304
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(join(3387..4178,4179..4247))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4010..4029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4248..4352)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttgacactc tatcattgat agagttattt tacctctccc tatcagtgat agagaaaact
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tggctagcag aggatcgcat
      121 caccatcacc atcacatcga agggcgcctt cttcacgagt tcggacttct cgagtcgccc
      181 aaggccctgg aagaggcgcc gtggcctcct ccagaggggg ctttcgtggg atttgtgctg
      241 agtcgaaaag aacccatgtg ggctgattta ctcgcgctgg ctgcggcaag agggggacgt
      301 gtccatcgtg ctcctgagcc ctataaagca ctgcgcgatc tcaaggaggc tcgtggcctc
      361 ctcgccaagg atctttctgt tttggccctg cgggagggat tggggcttcc tccgggcgac
      421 gatcccatgt tacttgcgta tttattggac cccagtaaca cgacacctga aggagtagcc
      481 cgtcgctatg ggggggaatg gaccgaagaa gcgggagaac gtgcggcact gtcggaacgc
      541 ctctttgcga acttatgggg ccgtcttgaa ggtgaagagc gtcttctgtg gctttaccgc
      601 gaagtggagc gtccgctgtc ggcagttctc gctcatatgg aagctacggg tgttagactt
      661 gacgtggcat acctgcgtgc gttgtcgtta gaagtggccg aagagattgc tagactcgaa
      721 gcagaagttt ttcggttagc gggacatcca tttaatctca attcccgtga ccagctggag
      781 cgcgttctct ttgatgaatt ggggctccct gccattggga aaaccgaaaa aaccggtaaa
      841 cgctcaacca gtgctgctgt gctggaggca ctgcgtgagg cccatccgat cgtggagaaa
      901 attctgcagt accgggagct cactaagttg aaaagcacat atatcgaagg cctgctgaaa
      961 gtagttcgcc cggacaccaa aaaagtccac acaagattta accagacggc gaccgcaacc
     1021 ggacgcctgt cctcatccga tccgaacttg cagaacatcc cggtccgtac tccgttgggt
     1081 cagcggatcc ggcgggcatt tatcgccggg gaaggttggc ttctggtcgc gctggactac
     1141 agccaaatcg aattgcgtgt gttagcacac ctttcgggag acgaaaattt gattcgggtg
     1201 tttcaagagg gtcgtgatat tcacacggaa accgcgagtt ggatgtttgg ggtgcctaga
     1261 gaggcagtgg atccacttat gcgtcgtgca gcaaaaacaa tcaatttcgg cgtcctctat
     1321 ggtatgagcg ctcaccgtct ttctcaggaa cttgccatcc cctacgaaga ggcccaggca
     1381 tttattgagc gttatttcca gagctttccg aaagttagag cctggattga gaaaaccttg
     1441 gaagaaggtc ggcgtcgggg gtatgtggag actctttttg gtcgtcggcg ctacgtaccg
     1501 gatctggaag ctcgtgtcaa gtctgtgcgc gaagctgccg aacgtatggc atttaatatg
     1561 ccggtccagg ggaccgcagc agatcttatg aagtcagcaa tggtcaaatt gttcccccgg
     1621 cttgaagaga tgggggcgcg tatgctgctt caggtacatg atgagcttgt cctggaagcc
     1681 ccgaaggaac gagcggaagc tgtggcccgt cttgctaaag aggtcatgga aggagtctac
     1741 ccactggccg tgccgttgga agtggaggtc ggcattggag aagattggtt atctgcgaaa
     1801 gaatgataat tgcctctgcc aaaattctgt cctcaagcgt tttagttcgt cggtcagttt
     1861 cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat
     1921 gaatgactgt cggccattcg atggtgtcgg gtcatgtgag caaaaggcca gcaaaaggcc
     1981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2701 tagatccttt taaactagtg aagttaccat cacggaaaaa ggttatgctg cttttaagac
     2761 ccactttcac atttaagttg tttttctaat ccgcatatga tcaattcaag gccgaataag
     2821 aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc
     2881 atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc
     2941 aatacgcaac ctaaagtaaa atgccccact gcgctgagtg catataatgc attctctagt
     3001 gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct
     3061 gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa agcacatcta
     3121 aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag
     3181 tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt
     3241 tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag tttacgggtt
     3301 gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt
     3361 ttatctaaac gagacatcat taattcctac caatgcttaa tcagtgaggc acctatctca
     3421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga gccacgctca
     3541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3601 cctgcaactt tatccgcctc catccagtct atcaattgtt gccgggaagc tagagtaagt
     3661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4321 atttagaaaa ataaacaaat aggggttccg cgcacatttc ccgggaactg ccagacatca
     4381 aataaaacaa aaggctcagt cggaagactg ggccttttgt tttatctgtt gtttgtcggt
     4441 gaacactctc ccgattgcga tacagaccct aatttcacat catatgacac taatcctaat
     4501 tttt
//
