LOCUS       sequence-310070-        7133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses His-tagged large fragment of Bst polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7133)
  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 7133)
  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(+)-Bst LF-6xHis
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     885..904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     988..1007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        988..1006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1007..1031
                     /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             1046..1068
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2852..2869
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2922..2940)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2936..2983
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3020..3475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3107..3126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3317..3338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3501..3605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3606..3674,3675..4466)
                     /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(3824..3843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4637..5225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5126..5145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5379..5396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5411..5550
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5476..5495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5636..5658
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5652..5843)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6652..7133,1..601))
                     /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 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      961 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     1021 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggc
     1081 ggagggagaa aagccgttgg aagaaatgga gtttgctatt gttgatgtaa ttaccgaaga
     1141 gatgctggca gataaagccg ctttagtggt tgaagtaatg gaggaaaatt accacgacgc
     1201 ccctatcgtt ggaattgcgt tggtcaatga acatgggcgc tttttcatgc gccctgagac
     1261 tgctcttgct gactcacaat ttcttgcttg gctggccgat gagactaaga agaaatctat
     1321 gttcgatgca aagcgtgccg ttgttgcttt gaagtggaaa ggcatcgaac ttcgcggtgt
     1381 agctttcgac ctgctgcttg cagcatacct tcttaatcca gctcaggatg cgggcgacat
     1441 cgcggctgta gcgaaaatga aacaatatga agctgttcgt tccgatgagg cagtctacgg
     1501 caagggagtc aaacgttctc ttccggatga acagacatta gccgagcatc tggtccgtaa
     1561 ggccgcagcc atctgggcac tggagcaacc tttcatggat gacctgcgca acaatgaaca
     1621 ggaccagctt ctgacaaagc tggaacagcc gttggcggca attttggcgg aaatggaatt
     1681 tactggcgtc aatgtggaca cgaagcgctt agaacaaatg ggatctgaac ttgccgagca
     1741 gttacgtgca atcgagcaac gtatttatga gctggcggga caggagttca atatcaatag
     1801 cccaaaacag ttgggggtaa tcctgtttga aaaactgcag ctgcccgtat tgaagaaaac
     1861 gaagaccggt tattcgacaa gcgctgatgt gttggaaaaa ttggcacccc accacgagat
     1921 cgtagagaat atcttgcatt accgccagtt gggcaagttg caaagcacgt atatcgaagg
     1981 ccttcttaag gtggtacgcc cagacacggg gaaagtccat acgatgttta accaagcttt
     2041 aacccaaact ggtcgcctta gtagtgcaga acccaattta caaaatattc ccatccgttt
     2101 agaagaaggc cgtaaaattc gtcaggcctt tgtgccatct gaacccgatt ggttgatttt
     2161 tgcagcggat tatagccaaa ttgagttacg cgtgttggca catatcgctg acgacgataa
     2221 tctgatcgaa gcatttcagc gcgatttgga catccatact aagacagcga tggacatctt
     2281 ccatgtaagt gaggaggaag tgacggcgaa catgcgccgc caggctaaag ctgttaactt
     2341 cggtatcgta tacggaatca gcgactacgg gttggctcag aacttgaaca ttacgcgtaa
     2401 ggaagcagcg gagtttattg agcgctactt cgcctctttc ccaggtgtta agcaatatat
     2461 ggaaaacatc gttcaggagg cgaaacaaaa ggggtatgtc acgacacttc ttcatcgccg
     2521 ccgttacctg cccgatatca ccagtcgtaa ctttaacgtg cgtagtttcg cggagcgcac
     2581 ggcgatgaat acccccattc agggttccgc agccgacatt atcaaaaagg cgatgattga
     2641 cttggccgcg cgcttaaagg aagagcaact tcaagcgcgc ctgttactgc aggtacatga
     2701 tgagcttatc ttagaggctc ccaaagagga gatcgaacgt ttgtgtgagt tggttcccga
     2761 agtcatggag caagctgtta cgttacgcgt accattaaag gtagactacc actacggtcc
     2821 tacatggtat gacgcgaaag gttcttccgg gcaccaccac caccaccact gagatccggc
     2881 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     2941 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     3001 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3061 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3361 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3421 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4141 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5341 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5401 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5461 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     5521 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     5581 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5641 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     5701 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     5761 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     5821 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     5881 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     5941 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     6001 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     6061 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     6121 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     6181 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     6241 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     6301 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     6361 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     6421 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     6481 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     6541 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     6601 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     6661 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6721 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     6781 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     6841 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     6901 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     6961 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     7021 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     7081 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gcc
//
