LOCUS       sequence-310074-        7596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses His-tagged HIV reverse transcriptase and untagged HIV 
            protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7596)
  AUTHORS   Kellner MJ, Ross JJ, Schnabl J, Dekens MPS, Matl M, Heinen R, 
            Grishkovskaya I, Bauer B, Stadlmann J, Menendez-Arias L, Straw AD, 
            Fritsche-Polanz R, Traugott M, Seitz T, Zoufaly A, Fodinger M, 
            Wenisch C, Zuber J, Pauli A, Brennecke J
  TITLE     A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay
            for Laboratory and Home Testing.
  JOURNAL   Front Mol Biosci. 2022 Apr 1;9:801309. doi: 
            10.3389/fmolb.2022.801309. eCollection 2022.
  PUBMED    35433827
REFERENCE   2  (bases 1 to 7596)
  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: "Front Mol 
            Biosci."; date: "2022-04-1"; volume: "9:801309. doi"; pages: " 
            10.3389/fmolb.2022.801309. eCollection 2022"
COMMENT     Sequence Label: pET21a(+)-HIV RT-6xHis-RBS-prot
FEATURES             Location/Qualifiers
     source          1..7596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     969..988
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1008..1085)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1291..1310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1394..1413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1394..1412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1413..1437
                     /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)."
     RBS             1452..1474
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3177..3194
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3791..3809)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3805..3852
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3889..4344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3976..3995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4186..4207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4370..4474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4475..4543,4544..5335)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4693..4712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6248..6265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6280..6419
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6345..6364)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6505..6527
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6521..6712)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7521..7596,1..1007))
                     /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"
ORIGIN
        1 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
       61 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      121 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      181 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
      241 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      301 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      361 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      421 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      481 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      541 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      601 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
      661 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
      721 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
      781 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
      841 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
      901 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
      961 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1021 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1081 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     1141 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     1201 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     1261 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     1321 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     1381 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     1441 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgccaatt agcccaattg
     1501 agactgtacc tgtaaagtta aaaccgggca tggacggccc taaggtgaag caatggccat
     1561 tgaccgagga gaaaatcaag gctttagtgg agatctgcac cgagatggaa aaagaaggta
     1621 aaatttccaa gattggaccg gaaaatccat acaacactcc cgtcttcgca attaagaaga
     1681 aggatagtac caagtggcgc aaacttgtgg acttccgcga gctgaacaaa cgcactcaag
     1741 acttctggga ggttcagctg ggaattcccc atcctgcagg attgaaaaag aagaaaagtg
     1801 ttaccgtgct tgacgtgggc gacgcatatt tctcagttcc cctggatgaa gacttccgca
     1861 aatacactgc gtttacgatc ccctcaatta ataacgaaac cccaggaatt cgctaccagt
     1921 ataatgtact gccgcaaggg tggaaaggaa gtccagccat ctttcaatca agcatgacga
     1981 aaattttaga accttttaaa aaacagaacc ccgatattgt tatttaccaa tacatggatg
     2041 acctgtatgt aggatcggac ttagagatcg gtcagcaccg tacaaagatt gaggagttgc
     2101 gccaacatct tcttcgttgg gggttaacaa cacccgataa aaagcatcaa aaagagccgc
     2161 catttttgtg gatgggttac gaattgcacc ctgataagtg gacggtgcag cctattgtgt
     2221 tgccggagaa ggatagttgg accgtaaatg acatccaaaa gctggtgggg aagcttaact
     2281 gggcgagcca aatttacccg gggattaagg tgcgccagct ttgcaaactg cttcgcggaa
     2341 ccaaagcact gactgaagtt atcccgctga cggaagaagc agagttggaa ctggccgaaa
     2401 accgtgaaat tttaaaagaa cccgtccacg gggtatatta tgatcccagt aaagacctga
     2461 ttgcagagat ccagaagcag ggacaagggc agtggacgta ccagatctat caagagcctt
     2521 tcaagaacct gaaaacgggc aaatatgcgc gtatgcgcgg tgcccacaca aacgacgtca
     2581 aacaacttac tgaggcggtt cagaaaatca ctaccgaaag tattgtcatt tggggaaaaa
     2641 ctccgaaatt caagttacca attcaaaaag agacatggga gacctggtgg actgagtatt
     2701 ggcaagctac atggatccca gagtgggagt ttgtgaatac tccccctttg gtgaagttat
     2761 ggtatcaact tgagaaggaa cctatcgtag gggctgaaac gttttacgtt gatggcgctg
     2821 caaatcgtga aacaaagctg ggtaaggcag gatatgtcac caataaaggt cgtcaaaagg
     2881 tagtgccgtt aacaaatacc acgaaccaga aaaccgaact tcaagccatc tatcttgccc
     2941 ttcaagactc cggtttagaa gtaaatatcg ttacggacag tcagtacgct ttaggcatta
     3001 ttcaagcgca accagataaa tctgagtcgg agcttgtgaa tcaaattatc gaacagttaa
     3061 tcaaaaagga aaaggtctac ttagcttggg tccctgcgca caagggcatc ggaggtaatg
     3121 aacaggttga taagttagtt agcgctggga tccgtaacat cttgggctcc ggctctcacc
     3181 accaccacca tcattgagaa ttcgctagca aaggaataag taatgcatca cgcccccgga
     3241 tccggggatc tggccttcct acaagggaag gccagggaat tttcttcaga gcagaccaga
     3301 gccaacagcc ccaccatttc ttcagagcag accagagcca acagccccac cagaagagag
     3361 cttcaggtct ggggtagaga caacaactcc ccctcagaag caggagccga tagacaagga
     3421 actgtatcct ttaacttccc tcagatcact ctatggcaac gacccctcgt cacaataaag
     3481 ataggggggc aactaaagga agctctatta gatacaggag cagatgatac agtattagaa
     3541 gaaatgagtt tgccaggaag atggaaacca aaaatgatag ggggaattgg aggttttatc
     3601 aaagtaagac agtatgatca gatactcata gaaatctgtg gacataaagc tataggtaca
     3661 gtattagtag gacctacacc tgtcaacata attggaagaa atctgttgac tcagattggt
     3721 tgcactttaa agcttagcta agatccggct gctaacaaag cccgaaagga agctgagttg
     3781 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     3841 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     3901 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3961 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4021 ttccccgtca agctctaaat cggggactcc ctttagggtt ccgatttagt gctttacggc
     4081 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4141 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4201 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4261 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4321 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4381 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4441 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4501 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4561 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4621 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4681 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4741 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4801 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4861 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4921 tttgcacaac atgggggacc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4981 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     5041 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5101 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5161 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5221 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5281 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5341 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6181 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6241 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     6301 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     6361 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     6421 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6481 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6541 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     6601 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     6661 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     6721 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     6781 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     6841 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     6901 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     6961 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     7021 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     7081 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     7141 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     7201 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     7261 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     7321 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     7381 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     7441 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     7501 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7561 tgcattaatg aatcggccaa cgcgcgggga gaggcg
//
