LOCUS       sequence-417252-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of DNA polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  AUTHORS   Bhadra S, Pothukuchy A, Shroff R, Cole AW, Byrom M, Ellefson JW, 
            Gollihar JD, Ellington AD
  TITLE     Cellular reagents for diagnostics and synthetic biology.
  JOURNAL   PLoS One. 2018 Aug 15;13(8):e0201681. doi: 
            10.1371/journal.pone.0201681. eCollection 2018.
  PUBMED    30110361
REFERENCE   2  (bases 1 to 5380)
  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: "2018-08-15"; volume: "13(8):e0201681. doi"; pages: " 
            10.1371/journal.pone.0201681. eCollection 2018"
COMMENT     Sequence Label: pATetO 6XHis-Phusion
FEATURES             Location/Qualifiers
     source          1..5380
                     /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(4777..5365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4857..4876)
                     /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 ggcgcatgat
     2041 tttagatgtg gattacataa ctgaagaagg aaaacctgtt attaggctat tcaaaaaaga
     2101 gaacggaaaa tttaagatag agcatgatag aacttttaga ccatacattt acgctcttct
     2161 cagggatgat tcaaagattg aagaagttaa gaaaataacg ggggaaaggc atggaaagat
     2221 tgtgagaatt gttgatgtag agaaggttga gaaaaagttt ctcggcaagc ctattaccgt
     2281 gtggaaactt tatttggaac atccccaaga tgttcccact attagagaaa aagttagaga
     2341 acatccagca gttgtggaca tcttcgaata cgatattcca tttgcaaaga gatacctcat
     2401 cgacaaaggc ctaataccaa tggaggggga agaagagcta aagattcttg ccttcgatat
     2461 agaaaccctc tatcacgaag gagaagagtt tggaaaaggc ccaattataa tgattagtta
     2521 tgcagatgaa aatgaagcaa aggtgattac ttggaaaaac atagatcttc catacgttga
     2581 ggttgtatca agcgagagag agatgataaa gagatttctc aggattatca gggagaagga
     2641 tcctgacatt atagttactt ataatggaga ctcattcgac ttcccatatt tagcgaaaag
     2701 ggcagaaaaa cttgggatta aattaaccat tggaagagat ggaagcgagc ccaagatgca
     2761 gagaataggc gatatgacgg ctgtagaagt caagggaaga atacatttcg acttgtatca
     2821 tgtaataaca aggacaataa atctcccaac atacacacta gaggctgtat atgaagcaat
     2881 ttttggaaag ccaaaggaga aggtatacgc cgacgagata gcaaaagcct gggaaagtgg
     2941 agagaacctt gagagagttg ccaaatactc gatggaagat gcaaaggcaa cttatgaact
     3001 cgggaaagaa ttccttccaa tggaaattca gctttcaaga ttagttggac aacctttatg
     3061 ggatgtttca aggtcaagca cagggaacct tgtagagtgg ttcttactta ggaaagccta
     3121 cgaaagaaac gaagtagctc caaacaagcc aagtgaagag gagtatcaaa gaaggctcag
     3181 ggagagctac acaggtggat tcgttaaaga gccagaaaag gggttgtggg aaaacatagt
     3241 atacctagat tttagagccc tatatccctc gattataatt acccacaatg tttctcccga
     3301 tactctaaat cttgagggat gcaagaacta tgatatcgct cctcaagtag gccacaagtt
     3361 ctgcaaggac atccctggtt ttataccaag tctcttggga catttgttag aggaaagaca
     3421 aaagattaag acaaaaatga aggaaactca agatcctata gaaaaaatac tccttgacta
     3481 tagacaaaaa gcgataaaac tcttagcaaa ttctttctac ggatattatg gctatgcaaa
     3541 agcaagatgg tactgtaagg agtgtgctga gagcgttact gcctggggaa gaaagtacat
     3601 cgagttagta tggaaggagc tcgaagaaaa gtttggattt aaagtcctct acattgacac
     3661 tgatggtctc tatgcaacta tcccaggagg agaaagtgag gaaataaaga aaaaggctct
     3721 agaatttgta aaatacataa attcaaagct ccctggactg ctagagcttg aatatgaagg
     3781 gttctataag aggggattct tcgttacgaa gaagaggtat gcagtaatag atgaagaagg
     3841 aaaagtcatt actcgtggtt tagagatagt taggagagat tggagtgaaa ttgcaaaaga
     3901 aactcaagct agagttttgg agacaatact aaaacacgga gatgttgaag aagctgtgag
     3961 aatagtaaaa gaagtaatac aaaagcttgc caattatgaa attccaccag agaagctcgc
     4021 aatatatgag cagataacaa gaccattaca tgagtataag gcgataggtc ctcacgtagc
     4081 tgttgcaaag aaactagctg ctaaaggagt taaaataaag ccaggaatgg taattggata
     4141 catagtactt agaggcgatg gtccaattag caatagggca attctagctg aggaatacga
     4201 tcccaaaaag cacaagtatg acgcagaata ttacattgag aaccaggttc ttccagcggt
     4261 acttaggata ttggagggat ttggatacag aaaggaagac ctcagatacc aaaagacaag
     4321 acaagtcggc ctaacttcct ggcttaacat taaaaaatcc ggtaccggcg gtggcggtgc
     4381 aaccgtaaag ttcaagtaca aaggcgaaga aaaagaggta gacatctcca agatcaagaa
     4441 agtatggcgt gtgggcaaga tgatctcctt cacctacgac gagggcggtg gcaagaccgg
     4501 ccgtggtgcg gtaagcgaaa aggacgcgcc gaaggagctg ctgcagatgc tggagaagca
     4561 gaaaaagtga taattgcctc tgccaaaatt ctgtcctcaa gcgttttagt tcgtcggtca
     4621 gtttcacctg atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa
     4681 agatgaatga ctgtcggcca ttcgatggtg tcgggtcatg tgagcaaaag gccagcaaaa
     4741 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5281 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5341 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
//
