LOCUS       sequence-332588-        5246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mutant HIV-2 RT p66 reverse transcriptase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  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 5246)
  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: pGV3341
FEATURES             Location/Qualifiers
     source          1..5246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        48..66
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    67..91
                     /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             1830..1874
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1912..1930)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1926..1973
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2144..2935)
                     /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(2936..3027)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4088..5170)
                     /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     5132..5151
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(5171..5246,1..2))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac
       61 tataggggaa ttgtgagcgg ataacaattc ccctgtagaa ataattttgt ttaactttaa
      121 taaggagata taccatggcg gtagcaaagg tcgagcccat taagatcacc cttaagccgg
      181 ggaaagatgg ccccaagttg cgtcaatggc cccttaccaa agagaagatc gaggctttaa
      241 aggaaatttg cgagaagatg gagaaagaag ggcagttgga agaagccccg ccaacgaatc
      301 cttataacac ccccacattc gctattaaga agaaggacaa gaacaagtgg cgtatgctga
      361 tcgacttccg cgaacttaac aaagtaactc aggattttac ggagatccag ctgggaattc
      421 cccatccagc tggtttggcg aagaagcgtc gtattacggt gcttgatgtt ggagacgcgt
      481 atttctccat cccgctgcat gaagacttcc gcccatacac cgcctttacg ctgccgtccg
      541 tcaacaacgc tgagcccggg aaacgttata tttacaaagt cttgccgcag gggtggaaag
      601 gtagtcctgc catctttcaa catacaatgc gccagattct tgaaccgttt cgtaaggcga
      661 acaaggacgt gattatcatt cagtacgtgg atgacattct tatcgcgagc gaccgcacag
      721 atcttgaaca tgatcgcgtt atcctgcaac ttaaagaatt gcttaataat ctgggcttct
      781 ccactcctga cgagaagttc cagaaagatc ctccttatca ctggatgggt tacgagctgt
      841 ggccaaccaa atggaagtta cagaagatcc agctgcccca gaaggaggtt tggaccgtaa
      901 acgacattca gaagttagta ggtgtattaa actgggccgc tcaattatat cctggtatta
      961 agacgaaaca tctttgtcgt ctgatccgcg gcaagttgac actgactgaa gaagttcaat
     1021 ggacagagct cgctgaagca gagcttgagg agaatcgcat catcttgtca caggaacaag
     1081 aggggcatta ctatcaggag gagaaggaac tggaagctac tgtgcagaag gaccaggaca
     1141 accaatggac gtacaagatt caccaggaag ataagatttt gaaggttgga aagtatgcca
     1201 aggtgaagaa tacgcacacc aatggtattc gcttgttggc gcaggttgtc cagaagatcg
     1261 gcaaggaagc ccttgtaatt tggggacgta tcccaaaatt tcatctgcct gttgaacgcg
     1321 atacttggga acagtggtgg gacaactact ggcaagtgac ttggataccc gactgggact
     1381 tcgtgtcgac acctcctctg gttcgtctgg ctttcaatct tgtaaaggac ccgattccgg
     1441 gtaccgagac attctatact gacgggagtt gcaatcgcca gtcaaaggaa ggcaaggcgg
     1501 gatatatcac cgaccgcgga aaggacaaag tgaagaaatt ggagcagacc acgaatcagc
     1561 aagccgaatt agaggcgttc gcaatggcat taactgatag cggacctaaa gtcaacatca
     1621 tcgttgatag tcagtacgtt atgggaattg tggcgtcgca acctaccgag tcagaaaata
     1681 agcttgtaaa ccagattatt gaagagatga caaagaagga agccatttat gtagcttggg
     1741 tcccggctca taaaggtatt ggaggaaatc aagaggttga tcatttagtt tcccagggca
     1801 ttcgtcaagt attataactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     1861 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     1921 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     1981 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     2041 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcatcgt
     2101 ggccggatct tgcggcccct cggcttgaac gaattgttag acattatttg ccgactacct
     2161 tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca
     2221 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact
     2281 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg
     2341 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc
     2401 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt
     2461 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc
     2521 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg
     2581 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc
     2641 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct
     2701 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat
     2761 caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat
     2821 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga
     2881 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatactct
     2941 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3001 ttgaatgtat ttagaaaaat aaacaaatag ctagctcact cggtcgctac gctccgggcg
     3061 tgagactgcg gcgggcgctg cggacacata caaagttacc cacagattcc gtggataagc
     3121 aggggactaa catgtgaggc aaaacagcag ggccgcgccg gtggcgtttt tccataggct
     3181 ccgccctcct gccagagttc acataaacag acgcttttcc ggtgcatctg tgggagccgt
     3241 gaggctcaac catgaatctg acagtacggg cgaaacccga caggacttaa agatccccac
     3301 cgtttccggc gggtcgctcc ctcttgcgct ctcctgttcc gaccctgccg tttaccggat
     3361 acctgttccg cctttctccc ttacgggaag tgtggcgctt tctcatagct cacacactgg
     3421 tatctcggct cggtgtaggt cgttcgctcc aagctgggct gtaagcaaga actccccgtt
     3481 cagcccgact gctgcgcctt atccggtaac tgttcacttg agtccaaccc ggaaaagcac
     3541 ggtaaaacgc cactggcagc agccattggt aactgggagt tcgcagagga tttgtttagc
     3601 taaacacgcg gttgctcttg aagtgtgcgc caaagtccgg ctacactgga aggacagatt
     3661 tggttgctgt gctctgcgaa agccagttac cacggttaag cagttcccca actgacttaa
     3721 ccttcgatca aaccacctcc ccaggtggtt ttttcgaagg ttaagtcagt tggggaactg
     3781 cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta
     3841 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga
     3901 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     3961 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4021 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4201 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4261 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4321 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     4381 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     4441 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4501 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     4561 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     4621 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     4681 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     4741 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     4801 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     4861 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     4921 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     4981 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5041 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5101 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     5161 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5221 gaaaggtttt gcgccattcg atggtg
//
