LOCUS       sequence-288122-        4621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the aTc-inducible expression of the large-fragment of 
            Bst DNAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4621)
  AUTHORS   Bhadra S, Riedel TE, Lakhotia S, Tran ND, Ellington AD
  TITLE     High-Surety Isothermal Amplification and Detection of SARS-CoV-2.
  JOURNAL   mSphere. 2021 May 19;6(3):e00911-20. doi: 10.1128/mSphere.00911-20.
  PUBMED    34011690
REFERENCE   2  (bases 1 to 4621)
  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: "10.1128/mSphere"; 
            journalName: "mSphere"; date: "2021-05-19- 19"; volume: "6"; issue: 
            "3"; pages: "e00911-20"
COMMENT     Sequence Label: pAtetO_6xHis-Bst-LF
FEATURES             Location/Qualifiers
     source          1..4621
                     /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(4018..4606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4098..4117)
                     /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 ggcgcgaaag
     2041 tcccagcagc gaggaagaaa aaccgctggc gaaaatggca tttactctgg cggatcgcgt
     2101 taccgaagaa atgctggcgg ataaagcggc cttggtagtg gaagtggtgg aagaaaacta
     2161 tcatgacgcg cctattgttg ggattgctgt tgttaacgag cacggacgct tctttctgcg
     2221 cccggaaacc gcgctggccg atcctcaatt cgtggcttgg ctgggagatg agaccaaaaa
     2281 aaaaagcatg ttcgattcta aacgcgcggc cgttgcgtta aaatggaaag gcatcgaact
     2341 gtgtggtgtg agctttgatc tcctgctggc tgcctacctc ctcgatccgg cccagggtgt
     2401 tgatgatgtg gccgcagcgg ccaaaatgaa gcaatatgag gccgtccgtc ccgatgaagc
     2461 tgtgtatggg aagggcgcca aacgcgcggt acctgatgaa ccagtcctgg cagagcacct
     2521 ggtccgcaaa gcggcagcta tttgggaact ggaacgcccc tttctggacg aattacgccg
     2581 taatgaacaa gatcgtcttc tggtagaact ggagcagccg ctgtcgtcaa ttctcgctga
     2641 aatggaattt gcgggtgtta aagttgacac gaaacgcctg gagcagatgg gtaaagaatt
     2701 ggccgaacag ctgggtaccg tcgaacaacg tatttatgag ttagcgggac aggaatttaa
     2761 tatcaattcc ccgaagcagc tgggggtgat tctctttgag aaattgcagc tccctgtcct
     2821 taagaaaacc aaaaccggtt actcaaccag tgctgatgtg ctggagaaac tggccccgta
     2881 ccatgagatc gtggagaata ttctgcacta ccggcagctc ggtaagttgc aaagcacata
     2941 tatcgaaggc ctgctgaaag tagttcgccc ggacaccaaa aaagtccaca caatctttaa
     3001 ccaggcgctg acccaaaccg gacgcctgtc ctcaaccgaa ccgaacttgc agaacatccc
     3061 gatccgtctt gaggagggtc ggaagatccg gcaggcattt gtccctagcg agagcgattg
     3121 gcttattttc gcggcagact acagccaaat cgaattgcgt gtgttagcac acattgcgga
     3181 agacgataat ttgatggagg cgtttcgccg cgatcttgat attcacacga aaaccgcgat
     3241 ggatattttt caggtgagtg aagatgaagt gactccaaac atgcgtcgtc aagcaaaagc
     3301 agtcaatttc ggcatcgtct atggtatcag cgattacggt cttgctcaga atcttaacat
     3361 cagccggaaa gaggccgccg aatttattga gcgttatttc gagagctttc cgggcgttaa
     3421 acgctacatg gagaacatcg tgcaagaagc taagcagaag gggtatgtga ccactcttct
     3481 gcatcgtcgg cgctacttac cggatattac tagtcgtaac ttcaatgtgc gctcatttgc
     3541 cgaacgtatg gcaatgaata ccccgatcca ggggtccgca gcagatatta tcaagaaagc
     3601 aatgatcgat ttgaacgccc ggcttaaaga ggaacgcctt caggcgcacc tgctgcttca
     3661 ggtacatgat gagcttatcc tggaagcccc gaaggaagaa atggaacgtc tgtgccgtct
     3721 tgttcctgag gtcatggaac aagccgtgac actgcgcgtg ccgttgaaag tggattacca
     3781 ctatggaagt acttggtacg atgcgaaatg ataattgcct ctgccaaaat tctgtcctca
     3841 agcgttttag ttcgtcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt
     3901 tttgcttttg gaggggcaga aagatgaatg actgtcggcc attcgatggt gtcgggtcat
     3961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4441 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4621 c
//
