LOCUS       sequence-356092-        4861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of MMLV RT in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  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 4861)
  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-MMLV RT
FEATURES             Location/Qualifiers
     source          1..4861
                     /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(4258..4846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4338..4357)
                     /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 ggcgcacttt
     2041 gaacatcgaa gatgagcacc gccttcacga gacctctaag gagccggacg tgtcacttgg
     2101 ctccacctgg ttatcggatt tccctcaagc ctgggccgaa acaggcggga tggggcttgc
     2161 ggtccgtcaa gcacctttaa ttatcccact taaagcaacg tcaactccgg tcagcatcaa
     2221 acagtacccg atgtcccaag aggcgcgctt agggattaag ccccatatcc aacgtcttct
     2281 tgaccaaggg atcttagttc cctgtcagtc gccgtggaat acgccactgc ttcctgtcaa
     2341 gaagccgggc accaacgact accgtcccgt ccaggatctg cgcgaggtta ataaacgtgt
     2401 tgaggacatc catccgacag tccctaaccc ttataacctt ctgagcggac ttccaccctc
     2461 tcaccagtgg tacactgtgt tggacttgaa agatgctttt ttttgtcttc gccttcatcc
     2521 gaccagtcag ccattgtttg cgttcgaatg gcgtgatccg gaaatgggta tttcgggaca
     2581 attaacatgg acccgcttac cccaaggttt taaaaacagt ccaactcttt ttgatgaggc
     2641 cctgcaccgt gacttggctg actttcgcat tcaacatccg gatcttattc ttttacagta
     2701 tgttgacgac cttcttttgg cagcaacttc cgaactggat tgccaacaag gcacacgcgc
     2761 ccttcttcag accttaggga atttagggta tcgtgcttcc gcaaaaaaag cgcaaatctg
     2821 ccagaaacaa gttaagtacc tgggctatct gctgaaagag gggcaacgtt ggttgactga
     2881 ggcacgcaag gaaaccgtca tgggacagcc tactcctaaa acgcctcgtc agcttcgcga
     2941 gttcttgggc actgcggggt tctgccgctt atggatcccg gggtttgcag aaatggccgc
     3001 gcctttatat cccttgacaa agactggaac tttatttaac tggggcccgg atcaacagaa
     3061 agcatatcaa gagattaaac aggcgctttt gacggctcca gctttaggtt tgccggatct
     3121 gaccaaacct tttgagttgt ttgttgacga aaaacagggt tatgccaaag gggttttgac
     3181 tcaaaagctt ggcccttggc gtcgccctgt cgcctatctt tctaaaaaac ttgatccagt
     3241 tgctgcaggt tggccgccct gtttacgtat ggtcgccgcc atcgccgtgt tgacgaaaga
     3301 cgctgggaaa ctgaccatgg gacagccgct ggtcatcttg gccccccatg ccgttgaggc
     3361 ccttgtcaag cagccgccgg accgctggct ttctaacgcg cgcatgacac actatcaggc
     3421 tctgctgctg gatactgacc gtgtgcaatt tgggccggtt gtcgcattaa acccagctac
     3481 attattaccc ttacctgaag agggattgca acacaattgc ctggatatct tagccgaggc
     3541 ccacggaact cgccccgacc tgactgatca gccccttcca gatgccgacc atacatggta
     3601 taccgatggt tcttcgcttc tgcaggaagg ccagcgtaag gcaggtgctg cagtcacgac
     3661 tgaaaccgaa gtcatttggg ccaaggcttt acctgcaggc acgagcgccc agcgtgcgga
     3721 gttgatcgcg ttaacccagg ctcttaaaat ggcggagggt aagaaactga atgtatacac
     3781 tgactcacgc tacgcatttg ccactgcaca cattcacggt gaaatttatc gtcgtcgcgg
     3841 gcttttaact tcggaaggaa aagagatcaa gaacaaggac gagattctgg ccttattgaa
     3901 ggcattattt ctgccgaaac gcctgtcaat tattcattgc cccggacatc aaaaaggaca
     3961 ttcggctgag gcacgcggga atcgtatggc tgaccaggcg gctcgcaaag cggccatcac
     4021 cgaaaccccg gacacttcga cgcttctttg ataattgcct ctgccaaaat tctgtcctca
     4081 agcgttttag ttcgtcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt
     4141 tttgcttttg gaggggcaga aagatgaatg actgtcggcc attcgatggt gtcgggtcat
     4201 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4261 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4321 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4381 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4441 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4501 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4561 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4621 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4681 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4741 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4801 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4861 c
//
