LOCUS       sequence-304729-        4504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the aTc-inducible expression of the Klentaq.
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-Klentaq
FEATURES             Location/Qualifiers
     source          1..4504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(138..764)
                     /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     669..688
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(join(771..1562,1563..1631))
                     /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     1394..1413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1632..1736)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1889..1944
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1895..1913
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1925..1943
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1958..1980
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2006..2023
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2024..2035
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(3901..4489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3981..4000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
       61 agattatcaa aaaggatctt cacctagatc cttttaaact agtgaagtta ccatcacgga
      121 aaaaggttat gctgctttta agacccactt tcacatttaa gttgtttttc taatccgcat
      181 atgatcaatt caaggccgaa taagaaggct ggctctgcac cttggtgatc aaataattcg
      241 atagcttgtc gtaataatgg cggcatacta tcagtagtag gtgtttccct ttcttcttta
      301 gcgacttgat gctcttgatc ttccaatacg caacctaaag taaaatgccc cactgcgctg
      361 agtgcatata atgcattctc tagtgaaaaa ccttgttggc ataaaaaggc taattgattt
      421 tcgagagttt catactgttt ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg
      481 cgatgactta gtaaagcaca tctaaaactt ttagcgttat tacgtaaaaa atcttgccag
      541 ctttcccctt ctaaagggca aaagtgagta tggtgcctat ctaacatctc aatggctaag
      601 gcgtcgagca aagcccgctt attttttaca tgccaataca atgtaggctg ctctacacct
      661 agcttctggg cgagtttacg ggttgttaaa ccttcgattc cgacctcatt aagcagctct
      721 aatgcgctgt taatcacttt acttttatct aaacgagaca tcattaattc ctaccaatgc
      781 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      841 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      901 atgataccgc gagagccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      961 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctatcaat
     1021 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1081 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1141 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1201 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1261 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1321 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1381 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1441 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1501 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1561 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1621 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1681 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1741 tttcccggga actgccagac atcaaataaa acaaaaggct cagtcggaag actgggcctt
     1801 ttgttttatc tgttgtttgt cggtgaacac tctcccgatt gcgatacaga ccctaatttc
     1861 acatcatatg acactaatcc taatttttgt tgacactcta tcattgatag agttatttta
     1921 cctctcccta tcagtgatag agaaaactag aaataatttt gtttaacttt aagaaggaga
     1981 tatacatatg gctagcagag gatcgcatca ccatcaccat cacatcgaag ggcgccttct
     2041 tcacgagttc ggacttctcg agtcgcccaa ggccctggaa gaggcgccgt ggcctcctcc
     2101 agagggggct ttcgtgggat ttgtgctgag tcgaaaagaa cccatgtggg ctgatttact
     2161 cgcgctggct gcggcaagag ggggacgtgt ccatcgtgct cctgagccct ataaagcact
     2221 gcgcgatctc aaggaggctc gtggcctcct cgccaaggat ctttctgttt tggccctgcg
     2281 ggagggattg gggcttcctc cgggcgacga tcccatgtta cttgcgtatt tattggaccc
     2341 cagtaacacg acacctgaag gagtagcccg tcgctatgga ggggaatgga ccgaagaagc
     2401 gggagaacgt gcggcactgt cggaacgcct ctttgcgaac ttatggggcc gtcttgaagg
     2461 tgaagagcgt cttctgtggc tttaccgcga agtggagcgt ccgctgtcgg cagttctcgc
     2521 tcatatggaa gctacgggtg ttagacttga cgtggcatac ctgcgtgcgt tgtcgttaga
     2581 agtggccgaa gagattgcta gactcgaagc agaagttttt cggttagcgg gacatccatt
     2641 taatctcaat tcccgtgacc agctggagcg cgttctcttt gatgaattgg ggctccctgc
     2701 cattgggaaa accgaaaaaa ccggtaaacg ctcaaccagt gctgctgtgc tggaggcact
     2761 gcgtgaggcc catccgatcg tggagaaaat tctgcagtac cgggagctca ctaagttgaa
     2821 aagcacatat atcgaccccc tgccggattt aattcacccg cgcaccggaa gactccacac
     2881 aagatttaac cagacggcga ccgcaaccgg acgcctgtcc tcatccgatc cgaacttgca
     2941 gaacatcccg gtccgtactc cgttgggtca gcggatccgg cgggcattta tcgccgagga
     3001 aggttggctt ctggtcgcgc tggactacag ccaaatcgaa ttgcgtgtgt tagcacacct
     3061 ttcgggagac gaaaatttga ttcgggtgtt tcaagagggt cgtgatattc acacggaaac
     3121 cgcgagttgg atgtttgggg tgcctcgaga ggcagtggat ccacttatgc gtcgtgcagc
     3181 aaaaacaatc aatttcggcg tcctctatgg tatgagcgct caccgtcttt ctcaggaact
     3241 tgccatcccc tacgaagagg cccaggcatt tattgagcgt tatttccaga gctttccgaa
     3301 agttagagcc tggattgaga aaaccttgga agaaggtcgg cgtcgggggt atgtggagac
     3361 tctttttggt cgtcggcgct acgtaccgga tctggaagct cgtgtcaagt ctgtgcgcga
     3421 agctgccgaa cgtatggcat ttaatatgcc ggtccagggg accgcagcag atcttatgaa
     3481 gttagcaatg gtcaaattgt tcccccggct tgaagagatg ggggcgcgta tgctgcttca
     3541 ggtacatgat gagcttgtcc tggaagcccc gaaggaacga gcggaagctg tggcccgtct
     3601 tgctaaagag gtcatggaag gagtctaccc actggccgtg ccgttggaag tggaggtcgg
     3661 cattggagaa gattggttat ctgcgaaaga atgataattg cctctgccaa aattctgtcc
     3721 tcaagcgttt tagttcgtcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg
     3781 gtttttgctt ttggaggggc agaaagatga atgactgtcg gccattcgat ggtgtcgggt
     3841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4321 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4501 gatc
//
