LOCUS       sequence-252336-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  AUTHORS   Lulla A, Hyvonen M
  TITLE     Protein expressionists toolkit
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5126)
  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            "unpublished"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(SS)
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11..29
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    29..53
                     /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             66..88
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             102..125
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             853..876
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(890..908)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(891..908)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(966..984)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      980..1027
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1263..1284)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1434..2516)
                     /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     2478..2497
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2517..2594)
                     /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     2733..2752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2906..2925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3025..3047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3085..3103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3171..3275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3276..3344,3345..4136)
                     /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(3494..3513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4307..4895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4796..4815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5049..5066
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgcagctgat taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa
       61 ataattttgt ttaactttaa gaaggagata tatccatgaa tcaccatcac catcaccatc
      121 accattctgg cactagtggc gatgacgcgg caattcaaca aacgttagcc aaaatgggca
      181 tcaaaagcag cgatattcag cccgcgcctg tagctggcat gaagacagtt ctgactaaca
      241 gcggcgtgtt gtacatcacc gatgatggta aacatatcat tcaggggcca atgtatgacg
      301 ttagtggcac ggctccggtc aatgtcacca ataagatgct gttaaagcag ttgaatgcgc
      361 ttgaaaaaga gatgatcgtt tataaagcgc cgcaggaaaa acacgtcatc accgtgttta
      421 ctgatattac ctctggttac tcccacaaac tgcatgagca aatggcagac tacaacgcgc
      481 tggggatcac cgtgcgttat cttgctttcc cgcgccaggg gctggacagc gatgcagaga
      541 aagaaatgaa agctatctgg tgtgcgaaag ataaaaacaa agcgtttgat gatgtgatgg
      601 caggtaaaag cgtcgcacca gccagttgcg acgtggatat tgccgaccat tacgcacttg
      661 gcgtccagct tggcgttagc ggtactccgg cagttgtgct gagcaatggc acacttgttc
      721 cgggttacca gccgccgaaa gagatgaaag aattcctcga cgaacaccaa aaaatgacca
      781 gcggtaaatc tggtacctcc ggttctactg aaaacctgta cttccagtga gaccttaatt
      841 aactcgagcg catggagcca cccgcagttc gaaaaataag cttgagtatt ctatagtgtc
      901 acctaaatcc cagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      961 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1021 ttttttgctg aaaggaggaa ctatatccgg ataacctggc gtaatagcga agaggcccgc
     1081 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta
     1141 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1201 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1261 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1321 aaaccgtcta tcagggcgat ggcccacagc ttgcatgcct gcaggtcgga agcataaagt
     1381 gtaaagcctg gggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     1441 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1501 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     1561 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     1621 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     1681 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     1741 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     1801 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     1861 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     1921 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     1981 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     2041 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     2101 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     2161 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     2221 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     2281 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     2341 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     2401 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     2461 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     2521 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac
     2581 cattcgatgg tgtcggcgag ctcgaattgt gaaccatcac cctaatcaag ttttttgggg
     2641 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2701 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2761 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2821 gcgccgctac agggcgcgtc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2881 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2941 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3001 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3061 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3121 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3181 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3241 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3301 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3361 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3421 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3481 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3541 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3601 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3661 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3721 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3781 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3841 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3901 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3961 ttgctgataa atctggagcc ggtgagcgtg gttctcgcgg tatcattgca gcactggggc
     4021 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4081 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4141 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4201 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4261 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4321 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4381 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4441 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4501 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4561 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4621 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4681 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4741 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4801 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4861 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4921 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4981 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5041 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5101 tccccgcgcg ttggccgatt cattaa
//
