LOCUS       sequence-213768-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 10-His DsbC TEV-peptide fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  AUTHORS   Buchko GW, Pulavarti SVSRK, Ovchinnikov V, Shaw EA, Rettie SA, Myler
            PJ, Karplus M, Szyperski T, Baker D, Bahl CD
  TITLE     Cytosolic expression, solution structures, and molecular dynamics 
            simulation of genetically encodable disulfide-rich de novo designed 
            peptides.
  JOURNAL   Protein Sci. 2018 Aug 26. doi: 10.1002/pro.3453.
  PUBMED    30152054
REFERENCE   2  (bases 1 to 5915)
  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: "Protein 
            Sci. 2018 Aug 26. doi: 10.1002/pro.3453."
COMMENT     Sequence Label: pCDB364
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        106..124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    129..145
                     /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             164..186
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             200..229
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(951..969)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      965..1012
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1049..1504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1136..1155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1346..1367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1596..2411)
                     /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(1596..2411)
                     /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     2319..2338
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2403..2422
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2533..3121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3022..3041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3275..3292
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3307..3446
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3372..3391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3532..3554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3548..3739)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4548..5630)
                     /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     5592..5611
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5631..5708)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5914..5915,1..18)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
       61 tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc gaaattaata cgactcacta
      121 tagggaaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
      181 aggagatata catatgggtc accatcatca ccaccatcac catcaccacg gtagcgcggc
      241 aattcaacaa acgttagcca aaatgggcat caaaagcagc gatattcagc ccgcgcccgt
      301 agctggcatg aagacagttc tgactaacag cggcgtgttg tacatcaccg atgatggtaa
      361 acatatcatt caggggccaa tgtatgacgt tagcggcacg gctccggtca atgtcaccaa
      421 taagatgctg ttaaagcagt tgaatgcgct tgaaaaagag atgatcgttt ataaagcgcc
      481 gcaggaaaaa cacgtcatca ccgtgtttac tgatattacc tgtggttact gccacaaact
      541 gcatgagcaa atggcagact ataacgcgct ggggatcacc gtgcgttatc ttgctttccc
      601 gcgccagggg ctggacagcg atgcagagaa agaaatgaaa gctatctggt gtgcgaaaga
      661 taaaaacaaa gcgtttgatg atgtgatggc aggtaaaagc gtcgcaccag ccagctgcga
      721 cgtggatatt gccgaccatt acgtacttgg cgtccagctt ggcgttagcg gtactccggc
      781 agttgtgctg agcaatggca cacttgttcc gggttaccag ccgccgaaag agatgaaaga
      841 attcctcgac gaacaccaaa aaatgaccag cggtaaaggt agcgaaaacc tgtattttca
      901 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
      961 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1021 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1081 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1141 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1201 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1261 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1321 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1381 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1441 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1501 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1561 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     1621 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1681 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1741 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1801 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1861 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1921 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1981 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2041 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2101 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2161 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2221 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2281 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2341 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2401 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2461 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2521 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2581 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2641 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2701 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2761 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2821 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2881 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2941 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3001 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3061 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3121 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3181 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3241 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3301 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     3361 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3421 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3481 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3541 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3601 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3661 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3721 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     3781 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     3841 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3901 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3961 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4021 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4081 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4141 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     4201 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     4261 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     4321 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     4381 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     4441 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     4501 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4561 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4621 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4681 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     4741 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4801 tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta
     4861 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4921 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4981 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     5041 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     5101 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     5161 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5221 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5281 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5341 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5401 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5461 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     5521 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5581 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5641 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     5701 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5761 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     5821 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5881 gagcccgaag tggcgagccc gatcttcccc atcgg
//
