LOCUS       sequence-217214-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as TEV cleavable 
            N-terminual 8His and DsbC (leaderless thiol:disulfide interchange 
            protein DsbC from E. coli) fusions with optional C-terminal 
            StrepII-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5114)
  AUTHORS   Lulla A, Hyvonen M
  TITLE     Protein expressionists toolkit
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5114)
  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            "unpublished"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: pExp-DsbC
FEATURES             Location/Qualifiers
     source          1..5114
                     /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    19..43
                     /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             56..78
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..115
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             831..854
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(868..886)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(869..886)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(944..962)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      958..1005
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1241..1262)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1412..2494)
                     /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     2456..2475
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2495..2572)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     2711..2730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2884..2903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3003..3025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3063..3081)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3149..3253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3254..3322,3323..4114)
                     /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     complement(3472..3491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4285..4873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4774..4793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5027..5044
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt
       61 ttaactttaa gaaggagata tatccatgaa tcaccatcac catcaccatc accattctgg
      121 cactagtggc gatgacgcgg caattcaaca aacgttagcc aaaatgggca tcaaaagcag
      181 cgatattcag cccgcgcctg tagctggcat gaagacagtt ctgactaaca gcggcgtgtt
      241 gtacatcacc gatgatggta aacatatcat tcaggggcca atgtatgacg ttagtggcac
      301 ggctccggtc aatgtcacca ataagatgct gttaaagcag ttgaatgcgc ttgaaaaaga
      361 gatgatcgtt tataaagcgc cgcaggaaaa acacgtcatc accgtgttta ctgatattac
      421 ctgtggttac tgccacaaac tgcatgagca aatggcagac tacaacgcgc tggggatcac
      481 cgtgcgttat cttgctttcc cgcgccaggg gctggacagc gatgcagaga aagaaatgaa
      541 agctatctgg tgtgcgaaag ataaaaacaa agcgtttgat gatgtgatgg caggtaaaag
      601 cgtcgcacca gccagttgcg acgtggatat tgccgaccat tacgcacttg gcgtccagct
      661 tggcgttagc ggtactccgg cagttgtgct gagcaatggc acacttgttc cgggttacca
      721 gccgccgaaa gagatgaaag aattcctcga cgaacaccaa aaaatgacca gcggtaaatc
      781 tggtaccgaa aacctgtact tccagtgaga ccttaattaa ctcgagcgca tggagccacc
      841 cgcagttcga aaaataagct tgagtattct atagtgtcac ctaaatccca gcttgatccg
      901 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
      961 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1021 atatccggat aacctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1081 tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1141 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1201 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1261 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1321 cccacagctt gcatgcctgc aggtcggaag cataaagtgt aaagcctggg gtgcctaatg
     1381 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1501 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     1561 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     1621 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     1681 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     1741 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     1801 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     1861 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     1921 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     1981 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     2041 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     2101 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     2161 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     2221 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     2281 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     2341 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     2401 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     2461 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     2521 ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcggcgagct
     2581 cgaattgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2641 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2701 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2761 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcct
     2821 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2881 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2941 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3001 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3061 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3121 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3181 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3241 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3301 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3361 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3421 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3481 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3541 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3601 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3661 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3721 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3781 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3841 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3901 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3961 tgagcgtggt tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4021 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4081 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4141 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4201 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4681 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4981 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5041 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5101 ttaatgcagc tgat
//
