LOCUS       sequence-332579-        6447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mutant HIV-2 RT p51 reverse transcriptase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6447)
  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 6447)
  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: pGV3340
FEATURES             Location/Qualifiers
     source          1..6447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             38..60
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1384..1401
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1414..1431)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1438..1455
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1474..1492)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1488..1535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1572..2027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1659..1678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1869..1890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2142..2933,2934..3002))
                     /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     2765..2784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3003..3095)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3176..3764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3665..3684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3918..3935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3950..4089
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4015..4034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4175..4197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4191..4382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4954..6036)
                     /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     5998..6017
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6037..6114)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6320..6339
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6427..6446
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6427..6445
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6446..6447,1..23)
                     /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)."
ORIGIN
        1 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
       61 tataccatgg cggtagcaaa ggtcgagccc attaagatca cccttaagcc ggggaaagat
      121 ggccccaagt tgcgtcaatg gccccttacc aaagagaaga tcgaggcttt aaaggaaatt
      181 tgcgagaaga tggagaaaga agggcagttg gaagaagccc cgccaacgaa tccttataac
      241 acccccacat tcgctattaa gaagaaggac aagaacaagt ggcgtatgct gatcgacttc
      301 cgcgaactta acaaagtaac tcaggatttt acggagatcc agctgggaat tccccatcca
      361 gctggtttgg cgaagaagcg tcgtattacg gtgcttgatg ttggagacgc gtatttctcc
      421 atcccgctgc atgaagactt ccgcccatac accgccttta cgctgccgtc cgtcaacaac
      481 gctgagcccg ggaaacgtta tatttacaaa gtcttgccgc aggggtggaa aggtagtcct
      541 gccatctttc aacatacaat gcgccagatt cttgaaccgt ttcgtaaggc gaacaaggac
      601 gtgattatca ttcagtacgt ggatgacatt cttatcgcga gcgaccgcac agatcttgaa
      661 catgatcgcg ttatcctgca acttaaagaa ttgcttaata atctgggctt ctccactcct
      721 gacgagaagt tccagaaaga tcctccttat cactggatgg gttacgagct gtggccaacc
      781 aaatggaagt tacagaagat ccagctgccc cagaaggagg tttggaccgt aaacgacatt
      841 cagaagttag taggtgtatt aaactgggcc gctcaattat atcctggtat taagacgaaa
      901 catctttgtc gtctgatccg cggcaagttg acactgactg aagaagttca atggacagag
      961 ctcgctgaag cagagcttga ggagaatcgc atcatcttgt cacaggaaca agaggggcat
     1021 tactatcagg aggagaagga actggaagct actgtgcaga aggaccagga caaccaatgg
     1081 acgtacaaga ttcaccagga agataagatt ttgaaggttg gaaagtatgc caaggtgaag
     1141 aatacgcaca ccaatggtat tcgcttgttg gcgcaggttg tccagaagat cggcaaggaa
     1201 gcccttgtaa tttggggacg tatcccaaaa tttcatctgc ctgttgaacg cgatacttgg
     1261 gaacagtggt gggacaacta ctggcaagtg acttggatac ccgactggga cttcgtgtcg
     1321 acacctcctc tggttcgtct ggctttcaat cttgtaaagg acccgattcc gggtaccctc
     1381 gagcatcacc atcaccatca ctaacctagg ttagtgatgg tgatggtgat gctcgagcat
     1441 caccatcacc atcactaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     1501 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     1561 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1621 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1921 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1981 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
     2041 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2101 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2161 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2221 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2281 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2341 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2401 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2461 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2521 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2581 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2641 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2701 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     2761 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2821 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2881 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2941 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3001 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
     3061 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
     3121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3301 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3841 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3901 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3961 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     4021 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4081 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4201 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4261 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4321 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4381 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4441 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4501 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4561 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4621 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4681 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4741 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4801 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac
     4861 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4921 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4981 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5041 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     5101 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     5161 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     5221 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     5281 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     5341 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     5401 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     5461 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5521 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     5581 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     5641 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     5701 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     5761 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5821 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5881 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5941 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     6001 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     6061 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     6121 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     6181 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     6241 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     6301 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     6361 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     6421 cgaaattaat acgactcact atagggg
//
