LOCUS       sequence-356095-        5344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Taq DNA polymerase in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5344)
  AUTHORS   Bhadra S, Nguyen V, Torres JA, Kar S, Fadanka S, Gandini C, Akligoh 
            H, Paik I, Maranhao AC, Molloy J, Ellington AD
  TITLE     Producing molecular biology reagents without purification.
  JOURNAL   PLoS One. 2021 Jun 1;16(6):e0252507. doi: 
            10.1371/journal.pone.0252507. eCollection 2021.
  PUBMED    34061896
REFERENCE   2  (bases 1 to 5344)
  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: "PLoS One.";
            date: "2021-06-1"; volume: "16(6):e0252507. doi"; pages: " 
            10.1371/journal.pone.0252507. eCollection 2021"
COMMENT     Sequence Label: pATetO 6XHis-Taq DNA Pol
FEATURES             Location/Qualifiers
     source          1..5344
                     /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
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRDNAKSFRCALLSHRDGAKVHLGT
                     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(4741..5329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4821..4840)
                     /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 cacgtaaaaa 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 ggcgcatgcg
     2041 tggtatgctg ccgctgttcg aaccgaaagg tcgtgttctg ctggttgacg gtcaccacct
     2101 ggcgtaccgt accttccacg cgctgaaagg tctgaccacc tctcgtggtg aaccggttca
     2161 ggcggtttac ggtttcgcga aatctctgct gaaagcgctg aaagaagacg gtgacgcggt
     2221 tatcgttgtt ttcgacgcga aagcgccgtc tttccgtcac gaagcgtacg gtggttacaa
     2281 agcgggtcgt gcgccgaccc cggaagactt cccgcgtcag ctggcgctga tcaaagaact
     2341 ggttgacctg ctgggtctgg cgcgtctgga agttccgggt tacgaagcgg acgacgttct
     2401 ggcgtctctg gcgaaaaaag cggaaaaaga aggttacgaa gttcgtatcc tgaccgcgga
     2461 caaagacctg taccagctgc tgtctgaccg tatccacgcg ctgcacccgg aaggttacct
     2521 gatcaccccg gcgtggctgt gggaaaaata cggtctgcgt ccggaccagt gggcggacta
     2581 ccgtgcgctg accggtgacg aatctgacaa cctgccgggt gttaaaggta tcggtgaaaa
     2641 aaccgcgcgt aaactgctgg aagaatgggg ttctctggaa gcgctgctga aaaacctgga
     2701 ccgtctgaaa ccggcgatcc gtgaaaaaat cctggcgcac atggacgacc tgaaactgtc
     2761 ttgggacctg gcgaaagttc gtaccgacct gccgctggaa gttgacttcg cgaaacgtcg
     2821 tgaaccggac cgtgaacgtc tgcgtgcgtt cctggaacgt ctggaattcg gttctctgct
     2881 gcacgaattc ggtctgctgg aatctccgaa agcgctggaa gaagcgccgt ggccgccgcc
     2941 ggaaggtgcg ttcgttggtt tcgttctgtc tcgtaaagaa ccgatgtggg cggacctgct
     3001 ggcgctggcg gcggcgcgtg gtggtcgtgt tcaccgtgcg ccggaaccgt acaaagcgct
     3061 gcgtgacctg aaagaagcgc gtggtctgct ggcgaaagac ctgtctgttc tggcgctgcg
     3121 tgaaggtctg ggtctgccgc cgggtgacga cccgatgctg ctggcgtacc tgctggaccc
     3181 gtctaacacc accccggaag gtgttgcgcg tcgttacggt ggtgaatgga ccgaagaagc
     3241 gggtgaacgt gcggcgctgt ctgaacgtct gttcgcgaac ctgtggggtc gtctggaagg
     3301 tgaagaacgt ctgctgtggc tgtaccgtga agttgaacgt ccgctgtctg cggttctggc
     3361 gcacatggaa gcgaccggtg ttcgtctgga cgttgcgtac ctgcgtgcgc tgtctctgga
     3421 agttgcggaa gaaatcgcgc gtctggaagc ggaagttttc cgtctggcgg gtcacccgtt
     3481 caacctgaac tctcgtgacc aactggaacg tgttctgttc gacgaactgg gtctgccggc
     3541 gatcggtaaa accgaaaaaa ccggtaaacg ttctacctct gcggcggttc tggaagcgct
     3601 gcgtgaagcg cacccgatcg ttgaaaaaat cctgcagtac cgtgaactga ccaaactgaa
     3661 atctacctac atcgacccgc tgccggacct gatccacccg cgtaccggtc gtctgcacac
     3721 ccgtttcaac cagaccgcga ccgcgaccgg tcgtctgtct tcttctgacc cgaacctgca
     3781 gaacatcccg gttcgtaccc cactgggtca gcgtatccgt cgtgcgttca tcgcggaaga
     3841 aggttggctt ctggttgcgc tggactactc tcagatcgaa ctgcgtgttc tggcgcacct
     3901 gtctggtgac gaaaacctga tccgtgtttt ccaggaaggt cgtgacatcc acaccgaaac
     3961 cgcgtcttgg atgttcggtg ttccgcgtga agcggttgac ccgctgatgc gtcgtgcggc
     4021 gaaaaccatc aacttcggtg ttctgtacgg tatgtctgcg caccgtctgt ctcaggaact
     4081 ggcgatcccg tacgaagaag cgcaggcgtt catcgaacgt tacttccagt ctttcccgaa
     4141 agttcgtgcg tggatcgaaa aaaccctgga agaaggtcgt cgtcgtggtt acgttgaaac
     4201 cctgttcggt cgtcgtcgtt acgttccgga cctggaagcg cgtgttaaat ctgttcgtga
     4261 agcggcggaa cgtatggcgt tcaacatgcc ggttcagggt accgcggcgg acctgatgaa
     4321 actggcgatg gttaaactgt tcccgcgtct ggaagaaatg ggtgcgcgta tgctgctgca
     4381 ggttcacgac gaactggttc tggaagcgcc gaaagaacgt gcggaagcgg ttgcgcgtct
     4441 ggcgaaagaa gttatggaag gtgtttaccc gctggcggtt ccgctggaag ttgaagttgg
     4501 tatcggtgaa gactggctgt ctgcgaaaga atgataattg cctctgccaa aattctgtcc
     4561 tcaagcgttt tagttcgtcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg
     4621 gtttttgctt ttggaggggc agaaagatga atgactgtcg gccattcgat ggtgtcgggt
     4681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5161 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5341 gatc
//
