LOCUS       sequence-228494-        3927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli expression plasmid of small tetraheme cytochrome (STC) from 
            Shewanella oneidensis MR-1 with cleavable Strep tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3927)
  AUTHORS   Huang J, Zarzycki J, Gunner MR, Parson WW, Kern JF, Yano J, Ducat 
            DC, Kramer DM
  TITLE     Mesoscopic to Macroscopic Electron Transfer by Hopping in a Crystal 
            Network of Cytochromes.
  JOURNAL   J Am Chem Soc. 2020 Jun 10;142(23):10459-10467. doi: 
            10.1021/jacs.0c02729. Epub 2020 May 29.
  PUBMED    32406683
REFERENCE   2  (bases 1 to 3927)
  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"; doi: 
            "10.1021/jacs.0c02729"; journalName: "J Am Chem Soc"; date: 
            "2020-06-10- 10"; volume: "142"; issue: "23"; pages: "10459-10467"
COMMENT     Sequence Label: pTSSX-STC
FEATURES             Location/Qualifiers
     source          1..3927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(196..201,202..217,218..224)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    232..248
                     /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)."
     CDS             352..375
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             376..387
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(686..704)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      700..747
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(977..1792)
                     /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(977..1792)
                     /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     1700..1719
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1793..1884)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1902..2537)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2735..3817)
                     /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     3779..3798
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3818..3895)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gcaagctgat ccgggcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
       61 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
      121 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
      181 ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg
      241 gataacaatt tcacaaagga aggagaattc atgagcaaaa aactattaag tgtgcttttc
      301 ggggcaagtc ttgcagcgtt agcgctgtcg ccaaccgcat ttgccgcgga ttggagccac
      361 ccgcagttcg aaaaaatcga agggcgcgcg gatcaaaaat tatcagactt ccatgctgag
      421 tcaggtggtt gtgagagctg tcataaagat ggtacccctt ctgctgacgg tgcctttgaa
      481 tttgcacaat gccaaagctg ccacggtaag ctttctgaaa tggacgcagt acacaagcca
      541 cacgatggca acttagtgtg cgctgactgc catgcggtac atgatatgaa tgtaggccaa
      601 aaacctactt gtgaaagctg ccatgacgat ggccgtactt ctgcgtctgt tctgaagaag
      661 tgataagtaa cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      721 tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc
      781 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
      841 cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg
      901 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      961 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1021 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1081 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1141 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1201 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     1261 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     1321 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat
     1381 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1441 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     1501 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     1561 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     1621 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     1681 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     1741 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc atactcttcc
     1801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1861 aatgtattta gaaaaataaa caaataggca tgctagcgca gaaacgtcct agaagatgcc
     1921 aggaggatac ttagcagaga gacaataagg ccggagcgaa gccgtttttc cataggctcc
     1981 gcccccctga cgaacatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg atggctccct cttgcgctct cctgttcccg
     2101 tcctgcggcg tccgtgttgt ggtggaggct ttacccaaat caccacgtcc cgttccgtgt
     2161 agacagttcg ctccaagctg ggctgtgtgc aagaaccccc cgttcagccc gactgctgcg
     2221 ccttatccgg taactatcat cttgagtcca acccggaaag acacgacaaa acgccactgg
     2281 cagcagccat tggtaactga gaattagtgg atttagatat cgagagtctt gaagtggtgg
     2341 cctaacagag gctacactga aaggacagta tttggtatct gcgctccact aaagccagtt
     2401 accaggttaa gcagttcccc aactgactta accttcgatc aaaccgcctc cccaggcggt
     2461 tttttcgttt acagagcagg agattacgac gatcgtaaaa ggatctcaag aagatccttt
     2521 acggattccc gacaccatca ctctagattt cagtgcaatt tatctcttca aatgtagcac
     2581 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     2641 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     2701 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2821 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2881 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2941 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3001 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3061 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3121 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3181 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     3241 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     3301 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     3361 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3421 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3481 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3541 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3601 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3661 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3721 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3781 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3841 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     3901 tctcgacgct ctcccttatg cgactcc
//
