LOCUS       sequence-356094-        4869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Taq DNA Ligase in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4869)
  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 4869)
  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 Ligase
FEATURES             Location/Qualifiers
     source          1..4869
                     /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"
     rep_origin      complement(4266..4854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4346..4365)
                     /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 cacatgaagg gcgcatgacc
     2041 ctggaggccg cgcgccgtcg cgtgaacgag ttacgcgatt taattcgtta tcacaactat
     2101 ctttactacg tgttggacgc ccctgaaatc tcagatgcag agtatgatcg cctgttacgc
     2161 gaacttaagg agctggaaga acgtttccct gagttacaga gccctgatag cccgactgaa
     2221 caagtagggg cacgcccgct tgaatccact ttccgccctg tgcgtcatcc gacacgtatg
     2281 tattctttag acaatgcgtt cagtttggat gaagtacgtg cgttcgaaga gcgcattgag
     2341 cgcgcgttgg gtcgtaaggg gcctttttta tataccgtgg agcataaggt tgatgggtta
     2401 tcagttaatt tgtattacga agaaggtatc ttagtatttg gcgcaacgcg cggcgatggg
     2461 gaaacgggtg aagaagtgac acaaaacctt cttactattc gcactatccc gcgtcgtctt
     2521 acgggagtac ctgatcgcct tgaagttcgt ggagaagtat acatgcccat cgaggcattt
     2581 cttcgtttaa accaagagtt agaagaagca ggcgaacgca ttttcaagaa cccgcgcaat
     2641 gccgcagccg gttcgcttcg tcaaaaggac ccccgtgtta ctgctcgtcg cggattgcgc
     2701 gcgacgttct acgccttggg gttgggtctt gaagagacgg ggcttaaatc gcagcatgac
     2761 cttttgttgt ggcttcgtga gcgtggtttc ccagtagaac acggatttac acgtgctttg
     2821 ggggcggaag gggtggagga agtttaccaa gcctggttaa aggagcgccg taagcttccc
     2881 ttcgaagcgg acggtgttgt cgtcaaattg gatgacctgg cactgtggcg tgagttgggg
     2941 tatacggcac gtgcaccacg tttcgcgtta gcctataagt ttcccgcaga ggagaaggag
     3001 acgcgtctgt tatcagtcgc tttccaagta ggacgtaccg ggcgtattac tccagtggga
     3061 gtacttgaac ccgtttttat cgaggggtca gaggtgtcgc gtgttacgtt acataacgag
     3121 agtttcattg aagagttgga tgtacgtatt ggggattggg tacttgtcca caaagctggc
     3181 ggcgtgatcc ctgaggtctt gcgcgtcctg aaagaacgcc gcaccgggga ggagaaacca
     3241 attttgtggc ctgaaaattg tcctgaatgt gggcatgccc tgatcaagga agggaaagta
     3301 catcgttgcc ctaacccact gtgccccgct aagcgtttcg aagcgattcg tcattacgcc
     3361 agccgcaaag ctatggatat ccaaggttta ggggagaaat taattgaaaa actgctggag
     3421 aaaggcttag tgcgtgatgt ggctgattta taccgtctta aaaaggagga cttggtaaac
     3481 ttggaacgta tgggcgagaa aagtgcggag aatttgttac gccagatcga ggagtccaaa
     3541 gggcgcgggc tggaacgcct gttatacgcc ttggggctgc ccggcgtggg tgaagtgctt
     3601 gcgcgtaatt tagccttacg cttcggccac atggaccgtc ttttagaggc cgggcttgag
     3661 gaccttcttg aggtggaagg tgtcggcgag ttgactgctc gcgcgattct gaatactctg
     3721 aaagaccctg agtttcgtga tttagttcgc cgtttgaaag aagcgggtgt cgaaatggaa
     3781 gctaaagaac gcgaaggtga agcattgaag ggtttgacgt tcgtcattac gggtgaactg
     3841 agccgcccgc gtgaggaagt caaagctctt ttgcgtcgct taggcgcaaa agtgaccgac
     3901 tccgtctctc gcaaaaccgg ttttttggta gtaggcgaga acccggggtc gaagttagaa
     3961 aaggcacgtg ctttaggagt ccccacactg tccgaagaag agctgtatcg ccttatcgag
     4021 gagcgtacag ggaaagatcc ccgtgcgctg acggcatgat aattgcctct gccaaaattc
     4081 tgtcctcaag cgttttagtt cgtcggtcag tttcacctga tttacgtaaa aacccgcttc
     4141 ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtcggccat tcgatggtgt
     4201 cgggtcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4861 cctttgatc
//
