LOCUS       sequence-338354-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His-tagged Bst-LF-Idaho in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  TITLE     pET-BstLF-Idaho
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7080)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET-Bst-LF-Idaho
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             121..144
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1945..1962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2015..2033)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2029..2076
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2113..2568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2200..2219)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2410..2431
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2660..3475)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2660..3475)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3383..3402
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3467..3486
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3597..4185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4086..4105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4339..4356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4371..4510
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4436..4455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4596..4618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4612..4803)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5612..6694)
                     /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     6656..6675
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6695..6772)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6978..6997
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca ctccgcgggt
      121 ctggaagttc tgttccaggg gcccatggaa agtcccagca gcgaggaaga aaaaccgctg
      181 gcgaaaatgg catttactct ggcggatcgc gttaccgaag aaatgctggc ggataaagcg
      241 gccttggtag tggaagtggt ggaagaaaac tatcatgacg cgcctattgt tgggattgct
      301 gttgttaacg agcacggacg cttctttctg cgcccggaaa ccgcgctggc cgatcctcaa
      361 ttcgtggctt ggctgggaga tgagaccaaa aaaaaaagca tgttcgattc taaacgcgcg
      421 gccgttgcgt taaaatggaa aggcatcgaa ctgtgtggtg tgagctttga tctcctgctg
      481 gctgcctacc tcctcgatcc ggcccagggt gttgatgatg tggccgcagc ggccaaaatg
      541 aagcaatatg aggccgtccg tcccgatgaa gctgtgtatg ggaagggcgc caaacgcgcg
      601 gtacctgatg aaccagtcct ggcagagcac ctggtccgca aagcggcagc tatttgggaa
      661 ctggaacgcc cctttctgga cgaattacgc cgtaatgaac aagatcgtct tctggtagaa
      721 ctggagcagc cgctgtcgtc aattctcgct gaaatggaat ttgcgggtgt taaagttgac
      781 acgaaacgcc tggagcagat gggtaaagaa ttggccgaac agctgggtac cgtcgaacaa
      841 cgtatttatg agttagcggg acaggaattt aatatcaatt ccccgaagca gctgggggtg
      901 attctctttg agaaattgca gctccctgtc cttaagaaaa ccaaaaccgg ttactcaacc
      961 agtgctgatg tgctggagaa actggccccg taccatgaga tcgtggagaa tattctgcac
     1021 taccggcagc tcggtaagtt gcaaagcaca tatatcgaag gcctgctgaa agtagttcgc
     1081 ccggacacca aaaaagtcca cacaatcttt aaccaggcgc tgacccaaac cggacgcctg
     1141 tcctcaaccg aaccgaactt gcagaacatc ccgatccgtc ttgaggaggg tcggaagatc
     1201 cggcaggcat ttgtccctag cgagagcgat tggcttattt tcgcggcaga ctacagccaa
     1261 atcgaattgc gtgtgttagc acacattgcg gaagacgata atttgatgga ggcgtttcgc
     1321 cgcgatcttg atattcacac gaaaaccgcg atggatattt ttcaggtgag tgaagatgaa
     1381 gtgactccaa acatgcgtcg tcaagcaaaa gcagtcaatt tcggcatcgt ctatggtatc
     1441 agcgattacg gtcttgctca gaatcttaac atcagccgga aagaggccgc cgaatttatt
     1501 gagcgttatt tcgagagctt tccgggcgtt aaacgctaca tggagaacat cgtgcaagaa
     1561 gctaagcaga aggggtatgt gaccactctt ctgcatcgtc ggcgctactt accggatatt
     1621 actagtcgta acttcaatgt gcgctcattt gccgaacgta tggcaatgaa taccccgatc
     1681 caggggtccg cagcagatat tatcaagaaa gcaatgatcg atttgaacgc ccggcttaaa
     1741 gaggaacgcc ttcaggcgca cctgctgctt caggtacatg atgagcttat cctggaagcc
     1801 ccgaaggaag aaatggaacg tctgtgccgt cttgttcctg aggtcatgga acaagccgtg
     1861 acactgcgcg tgccgttgaa agtggattac cactatggaa gtacttggta cgatgcgaaa
     1921 taaaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
     1981 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2041 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     2101 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2161 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2221 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2281 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2341 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2401 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2461 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2521 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2581 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2641 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     2701 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     2761 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     2821 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     2881 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     2941 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     3001 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     3061 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     3121 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     3181 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     3241 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     3301 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     3361 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     3421 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     3481 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     3541 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3601 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3661 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3721 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3781 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3841 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3901 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3961 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4021 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4081 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4141 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4201 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4261 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4321 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4381 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     4441 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4501 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4561 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4621 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4681 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4741 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4801 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     4861 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4921 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4981 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     5041 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     5101 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     5161 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     5221 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     5281 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     5341 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     5401 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     5461 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     5521 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     5581 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5641 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5701 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     5761 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     5821 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     5881 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     5941 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     6001 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     6061 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     6121 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     6181 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     6241 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     6301 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     6361 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     6421 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     6481 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     6541 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     6601 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     6661 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     6721 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     6781 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     6841 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     6901 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     6961 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     7021 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
