LOCUS       sequence-297110-        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"
     promoter        1..56
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7..25
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             118..135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             136..147
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(2130..2718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2210..2229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2871..3497)
                     /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     3402..3421
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(join(3504..4295,4296..4364))
                     /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     4127..4146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4365..4469)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttgacactc tatcattgat agagttattt tacctctccc tatcagtgat agagaaaact
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tggctagcag aggatcgcat
      121 caccatcacc atcacatcga agggcgcgaa agtcccagca gcgaggaaga aaaaccgctg
      181 gcgaaaatgg catttactct ggcggatcgc gttaccgaag aaatgctggc ggataaagcg
      241 gccttggtag tggaagtggt ggaagaaaac tatcatgacg cgcctattgt tgggattgct
      301 gttgttaacg agcacggacg cttctttctg cgcccggaaa ccgcgctggc cgatcctcaa
      361 ttcgtggctt ggctgggaga tgagaccaaa aaaaaaagca tgttcgattc taaacgcgcg
      421 gccgttgcgt taaaatggaa aggcatcgaa ctgtgtggtg tgagctttga tctcctgctg
      481 gctgcctacc tcctcgatcc ggcccagggt gttgatgatg tggccgcagc ggccaaaatg
      541 aagcaatatg aggccgtccg tcccgatgaa gctgtgtatg ggaagggcgc caaacgcgcg
      601 gtacctgatg aaccagtcct ggcagagcac ctggtccgca aagcggcagc tatttgggaa
      661 ctggaacgcc cctttctgga cgaattacgc cgtaatgaac aagatcgtct tctggtagaa
      721 ctggagcagc cgctgtcgtc aattctcgct gaaatggaat ttgcgggtgt taaagttgac
      781 acgaaacgcc tggagcagat gggtaaagaa ttggccgaac agctgggtac cgtcgaacaa
      841 cgtatttatg agttagcggg acaggaattt aatatcaatt ccccgaagca gctgggggtg
      901 attctctttg agaaattgca gctccctgtc cttaagaaaa ccaaaaccgg ttactcaacc
      961 agtgctgatg tgctggagaa actggccccg taccatgaga tcgtggagaa tattctgcac
     1021 taccggcagc tcggtaagtt gcaaagcaca tatatcgaag gcctgctgaa agtagttcgc
     1081 ccggacacca aaaaagtcca cacaatcttt aaccaggcgc tgacccaaac cggacgcctg
     1141 tcctcaaccg aaccgaactt gcagaacatc ccgatccgtc ttgaggaggg tcggaagatc
     1201 cggcaggcat ttgtccctag cgagagcgat tggcttattt tcgcggcaga ctacagccaa
     1261 atcgaattgc gtgtgttagc acacattgcg gaagacgata atttgatgga ggcgtttcgc
     1321 cgcgatcttg atattcacac gaaaaccgcg atggatattt ttcaggtgag tgaagatgaa
     1381 gtgactccaa acatgcgtcg tcaagcaaaa gcagtcaatt tcggcatcgt ctatggtatc
     1441 agcgattacg gtcttgctca gaatcttaac atcagccgga aagaggccgc cgaatttatt
     1501 gagcgttatt tcgagagctt tccgggcgtt aaacgctaca tggagaacat cgtgcaagaa
     1561 gctaagcaga aggggtatgt gaccactctt ctgcatcgtc ggcgctactt accggatatt
     1621 actagtcgta acttcaatgt gcgctcattt gccgaacgta tggcaatgaa taccccgatc
     1681 caggggtccg cagcagatat tatcaagaaa gcaatgatcg atttgaacgc ccggcttaaa
     1741 gaggaacgcc ttcaggcgca cctgctgctt caggtacatg atgagcttat cctggaagcc
     1801 ccgaaggaag aaatggaacg tctgtgccgt cttgttcctg aggtcatgga acaagccgtg
     1861 acactgcgcg tgccgttgaa agtggattac cactatggaa gtacttggta cgatgcgaaa
     1921 tgataattgc ctctgccaaa attctgtcct caagcgtttt agttcgtcgg tcagtttcac
     1981 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
     2041 tgactgtcgg ccattcgatg gtgtcgggtc atgtgagcaa aaggccagca aaaggccagg
     2101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2821 atccttttaa actagtgaag ttaccatcac ggaaaaaggt tatgctgctt ttaagaccca
     2881 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
     2941 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
     3001 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
     3061 acgcaaccta aagtaaaatg ccccactgcg ctgagtgcat ataatgcatt ctctagtgaa
     3121 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
     3181 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
     3241 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
     3301 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
     3361 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
     3421 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
     3481 tctaaacgag acatcattaa ttcctaccaa tgcttaatca gtgaggcacc tatctcagcg
     3541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagagcc acgctcaccg
     3661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3721 gcaactttat ccgcctccat ccagtctatc aattgttgcc gggaagctag agtaagtagt
     3781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4441 tagaaaaata aacaaatagg ggttccgcgc acatttcccg ggaactgcca gacatcaaat
     4501 aaaacaaaag gctcagtcgg aagactgggc cttttgtttt atctgttgtt tgtcggtgaa
     4561 cactctcccg attgcgatac agaccctaat ttcacatcat atgacactaa tcctaatttt
     4621 t
//
