LOCUS       sequence-330566-        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  JOURNAL   Unpublished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            "Unpublished"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     complement(61..80)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     271..292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(521..1336)
                     /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(521..1336)
                     /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     1244..1263
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1328..1347
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1458..2046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1947..1966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2200..2217
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2232..2371
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2297..2316)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2457..2479
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2473..2664)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3473..4555)
                     /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     4517..4536
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4556..4633)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4839..4858
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4942..4961
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4942..4960
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4961..4985
                     /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             5000..5022
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5035..5052
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5062..5085
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6886..6903
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6956..6974)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6970..7017
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(7054..7080,1..429)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
       61 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      121 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      181 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      241 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      301 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      361 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
      421 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      481 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
      541 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
      601 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
      661 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
      721 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
      781 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
      841 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
      901 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
      961 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     1021 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     1081 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     1141 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     1201 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     1261 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     1321 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     1381 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1441 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1501 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1561 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1621 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1681 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1741 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1801 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1861 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1921 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1981 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2041 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2101 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2161 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2221 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2281 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     2341 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     2401 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     2461 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     2521 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     2581 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     2641 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     2701 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     2761 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     2821 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     2881 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     2941 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     3001 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     3061 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     3121 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     3181 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     3241 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     3301 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     3361 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3421 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     3481 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3541 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     3601 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3661 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     3721 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     3781 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     3841 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     3901 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     3961 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     4021 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     4081 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     4141 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     4201 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     4261 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     4321 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     4381 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     4441 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4501 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     4561 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4621 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     4681 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     4741 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     4801 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     4861 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     4921 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     4981 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat gaaacatcac
     5041 catcaccatc actccgcggg tctggaagtt ctgttccagg ggcccatgga aagtcccagc
     5101 agcgaggaag aaaaaccgct ggcgaaaatg gcatttactc tggcggatcg cgttaccgaa
     5161 gaaatgctgg cggataaagc ggccttggta gtggaagtgg tggaagaaaa ctatcatgac
     5221 gcgcctattg ttgggattgc tgttgttaac gagcacggac gcttctttct gcgcccggaa
     5281 accgcgctgg ccgatcctca attcgtggct tggctgggag atgagaccaa aaaaaaaagc
     5341 atgttcgatt ctaaacgcgc ggccgttgcg ttaaaatgga aaggcatcga actgtgtggt
     5401 gtgagctttg atctcctgct ggctgcctac ctcctcgatc cggcccaggg tgttgatgat
     5461 gtggccgcag cggccaaaat gaagcaatat gaggccgtcc gtcccgatga agctgtgtat
     5521 gggaagggcg ccaaacgcgc ggtacctgat gaaccagtcc tggcagagca cctggtccgc
     5581 aaagcggcag ctatttggga actggaacgc ccctttctgg acgaattacg ccgtaatgaa
     5641 caagatcgtc ttctggtaga actggagcag ccgctgtcgt caattctcgc tgaaatggaa
     5701 tttgcgggtg ttaaagttga cacgaaacgc ctggagcaga tgggtaaaga attggccgaa
     5761 cagctgggta ccgtcgaaca acgtatttat gagttagcgg gacaggaatt taatatcaat
     5821 tccccgaagc agctgggggt gattctcttt gagaaattgc agctccctgt ccttaagaaa
     5881 accaaaaccg gttactcaac cagtgctgat gtgctggaga aactggcccc gtaccatgag
     5941 atcgtggaga atattctgca ctaccggcag ctcggtaagt tgcaaagcac atatatcgaa
     6001 ggcctgctga aagtagttcg cccggacacc aaaaaagtcc acacaatctt taaccaggcg
     6061 ctgacccaaa ccggacgcct gtcctcaacc gaaccgaact tgcagaacat cccgatccgt
     6121 cttgaggagg gtcggaagat ccggcaggca tttgtcccta gcgagagcga ttggcttatt
     6181 ttcgcggcag actacagcca aatcgaattg cgtgtgttag cacacattgc ggaagacgat
     6241 aatttgatgg aggcgtttcg ccgcgatctt gatattcaca cgaaaaccgc gatggatatt
     6301 tttcaggtga gtgaagatga agtgactcca aacatgcgtc gtcaagcaaa agcagtcaat
     6361 ttcggcatcg tctatggtat cagcgattac ggtcttgctc agaatcttaa catcagccgg
     6421 aaagaggccg ccgaatttat tgagcgttat ttcgagagct ttccgggcgt taaacgctac
     6481 atggagaaca tcgtgcaaga agctaagcag aaggggtatg tgaccactct tctgcatcgt
     6541 cggcgctact taccggatat tactagtcgt aacttcaatg tgcgctcatt tgccgaacgt
     6601 atggcaatga ataccccgat ccaggggtcc gcagcagata ttatcaagaa agcaatgatc
     6661 gatttgaacg cccggcttaa agaggaacgc cttcaggcgc acctgctgct tcaggtacat
     6721 gatgagctta tcctggaagc cccgaaggaa gaaatggaac gtctgtgccg tcttgttcct
     6781 gaggtcatgg aacaagccgt gacactgcgc gtgccgttga aagtggatta ccactatgga
     6841 agtacttggt acgatgcgaa ataaaagctt gcggccgcac tcgagcacca ccaccaccac
     6901 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     6961 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     7021 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
//
