LOCUS       sequence-332586-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mutant Bst polymerase (Bst-LF D720A) for loop mediated
            isothermal amplification (LAMP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  AUTHORS   Sherrill-Mix S, Hwang Y, Roche AM, Glascock A, Weiss SR, Li Y, 
            Haddad L, Deraska P, Monahan C, Kromer A, Graham-Wooten J, Taylor 
            LJ, Abella BS, Ganguly A, Collman RG, Van Duyne GD, Bushman FD
  TITLE     Detection of SARS-CoV-2 RNA using RT-LAMP and molecular beacons.
  JOURNAL   Genome Biol. 2021 Jun 3;22(1):169. doi: 10.1186/s13059-021-02387-y.
  PUBMED    34082799
REFERENCE   2  (bases 1 to 5378)
  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: "Genome 
            Biol."; date: "2021-06-3"; volume: "22(1):169. doi"; pages: " 
            10.1186/s13059-021-02387-y"
COMMENT     Sequence Label: pGV3319
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..48
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        29..47
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    48..72
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             128..145
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1943..1987
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2025..2043)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2039..2086
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2257..3048)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3049..3140)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4201..5283)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5245..5264
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5284..5361)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cccttatgcg actcctgcat taggaaatta atacgactca ctatagggga attgtgagcg
       61 gataacaatt cccctgtaga aataattttg tttaacttta ataaggagat ataccatggg
      121 cagcagccat caccatcatc accacagcca ggatccgtca tcagaagagg aaaaaccgct
      181 tgccaagatg gcatttacgc ttgctgaccg cgtgacggag gagatgcttg ccgacaaggc
      241 ggcgcttgtc gttgaagtgg tcgaggaaaa ttatcatgat gcgccgatcg tcggcatcgc
      301 tgtggtcaac gaacatggac ggtttttcct gcgcccggag acggcgcttg ccgatccgca
      361 gtttgtcgcc tggcttggtg atgaaacgaa gaaaaaaagc atgtttgact caaagcgcgc
      421 ggcagtcgcc ttgaaatgga aaggaattga gctatgcggc gtttcctttg atttattgct
      481 ggccgcctat ttgcttgatc cggcgcaagg tgttgatgat gtggctgccg cagcaaaaat
      541 gaagcaatac gaagcggtgc gcccggatga agcggtgtat ggcaaagggg cgaagcgggc
      601 cgtgccggat gagccagtgc tcgccgagca tttggtccgc aaggcggcgg cgatttgggc
      661 gctcgaacgg ccgtttttgg atgagctgcg ccgcaacgaa caagatcggt tgctcgtcga
      721 gctcgagcag ccgttgtctt cgattttggc ggaaatggaa tttgccggag tgaaagtgga
      781 tacgaagcgg ctcgaacaga tgggcgaaga gctcgccgag cagctgcgca cggtcgagca
      841 gcgcatttat gagctcgccg gccaagaatt caacatcaat tcaccgaaac agctcggcgt
      901 cattttattt gaaaaactgc agctgcccgt cttgaaaaaa acgaaaaccg gctactccac
      961 ttcggcggat gtgcttgaaa aacttgcgcc ttatcacgag atcgtggaaa acattttgca
     1021 ttaccgccag cttggcaagt tgcagtcgac gtatattgaa ggattgctga aagtcgtgcg
     1081 acccgataca aagaaggtgc atacgatttt caatcaggcg ttgacgcaaa ccggacggct
     1141 cagctcgacg gagccgaact tgcaaaacat tccgattcgg cttgaggaag gacggaaaat
     1201 ccgccaagcg ttcgtgccat cggagtctga ttggctcatt ttcgccgccg actactcgca
     1261 aattgagttg cgcgtcctcg cccatattgc ggaagatgac aatttaatgg aagcgttccg
     1321 ccgcgatttg gatatccata cgaaaacagc gatggacatt ttccaagtga gcgaggacga
     1381 agtgacgccc aacatgcgcc gtcaggcgaa ggcggtcaac tttgggatcg tttacgggat
     1441 cagtgcttac ggcttggcgc aaaacttaaa tatttcgcgc aaagaggcgg ctgaattcat
     1501 cgagcgctac ttcgaaagct tccctggcgt gaagcggtat atggaaaaca ttgtgcaaga
     1561 agcaaaacag aaagggtatg tgacgacgct gctgcatcgg cgccgctatt tgccggatat
     1621 cacgagccgc aacttcaacg tccgcagctt tgctgaacgg atggcgatga acacgccgat
     1681 tcaagggagc gccgctgaca ttattaaaaa ggcgatgatc gatctgaacg ccagactgaa
     1741 ggaagagcgg ctgcaagcgc gccttttgct gcaggtgcat gacgagctca ttttggaggc
     1801 gccgaaagaa gagatggagc ggctgtgccg gctcgttccg gaagtgatgg agcaagcggt
     1861 cacacttcgc gtgccgctca aagtcgatta ccattatggc tcgacgtggt atgatgcgaa
     1921 ataaggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
     1981 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
     2041 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     2101 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     2161 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat cgtggccgga
     2221 tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta ccttggtgat
     2281 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     2341 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     2401 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     2461 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     2521 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     2581 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     2641 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     2701 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     2761 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     2821 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     2881 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     2941 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     3001 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac tcttcctttt
     3061 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3121 tatttagaaa aataaacaaa tagctagctc actcggtcgc tacgctccgg gcgtgagact
     3181 gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata agcaggggac
     3241 taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag gctccgccct
     3301 cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc
     3361 aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc caccgtttcc
     3421 ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg gatacctgtt
     3481 ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac tggtatctcg
     3541 gctcggtgta ggtcgttcgc tccaagctgg gctgtaagca agaactcccc gttcagcccg
     3601 actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag cacggtaaaa
     3661 cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt agctaaacac
     3721 gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag atttggttgc
     3781 tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact taaccttcga
     3841 tcaaaccacc tccccaggtg gttttttcga aggttaagtc agttggggaa ctgcttaacc
     3901 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag
     3961 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt
     4021 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     4081 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     4141 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     4201 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4261 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4321 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4381 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4441 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     4501 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     4561 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     4621 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     4681 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     4741 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     4801 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     4861 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     4921 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     4981 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     5041 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     5101 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     5161 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     5221 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     5281 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5341 tttgcgccat tcgatggtgt ccgggatctc gacgctct
//
