LOCUS       sequence-349901-        6527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression plasmid expressing Human PDI and Yeast Erv1p to aid the
            expression of disulfide-rich proteins in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6527)
  AUTHORS   Yu DS, Outram MA, Crean E, Smith A, Sung Y, Darma R, Sun X, Ma L, 
            Jones DA, Solomon PS, Williams SJ
  TITLE     Optimised production of disulfide-bonded fungal effectors in E. coli
            using CyDisCo and FunCyDisCo co-expression approaches
  JOURNAL   bioRxiv 2021.08.31.458447
REFERENCE   2  (bases 1 to 6527)
  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: "bioRxiv 
            2021.08.31.458447"
COMMENT     Sequence Label: pACYC-T7CyDisCo
FEATURES             Location/Qualifiers
     source          1..6527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(144..149,150..166,167..172))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      284..829
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     730..749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     946..963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1344..1446
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1447..2106
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1513..1532)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3959..3978
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4006..4024
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4025..4049
                     /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             4064..4086
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      4680..4751
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4767..4794
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4823..4841
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4842..4866
                     /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             4881..4903
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      6406..6477
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6493..6520
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 gttccggcta gctgcagtca taagttaggt acctagctgc agtcataagg gacttagggt
       61 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
      121 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
      181 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
      241 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
      301 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
      361 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
      421 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
      481 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
      541 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
      601 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
      661 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
      721 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
      781 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
      841 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
      901 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
      961 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     1021 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     1081 tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt
     1141 agctgaacag gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca
     1201 tcatacacta aatcagtaag ttggcagcat cacccgacgc actttgcgcc gaataaatac
     1261 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
     1321 agccctgggc caaaaaaaga cgttgatcgg cacgtaagag gttccaactt tcaccataat
     1381 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     1441 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1501 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1561 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1621 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     1681 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1741 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1801 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1861 attgagaata tgtttttcgt gtcagccaat ccctgggtga gtttcaccag ttttgattta
     1921 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1981 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     2041 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     2101 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggttgcta cgcctgaata
     2161 agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt
     2221 cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt ttattgacta
     2281 ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct caggctctcc
     2341 ccgtggaggt aataattgac gatatgatca tttattctgc ctcccagagc ctgataaaaa
     2401 cggttagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     2461 gatgcagatc cggaacataa tggtgcaggg cgcttgtttc ggcgtgggta tggtggcagg
     2521 ccccgtggcc gggggactgt tgggcgctgc cggcacctgt cctacgagtt gcatgataaa
     2581 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctacc
     2641 ggcagcggtg cggactgttg taactcagaa taagaaatga ggccgctcat ggcgttgact
     2701 ctcagtcata gtatcgtggt atcaccggtt ggttccactc tctgttgcgg gcaacttcag
     2761 cagcacgtag gggacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     2821 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     2881 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     2941 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc
     3001 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     3061 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     3121 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     3181 cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc
     3241 tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg
     3301 taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     3361 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     3421 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     3481 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     3541 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     3601 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     3661 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     3721 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgacgctct cccttatgcg
     3781 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     3841 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     3901 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     3961 tgatgtcggc gatataggca tctctatatc tctatcccgc gaaattaata cgactcacta
     4021 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     4081 aggagatata ccatatgaag gctattgaca agatgacgga caacccgccg caagaagggt
     4141 tatcaggtcg taaaatcatt tacgacgagg acggcaagcc ttgtcgttct tgtaatactc
     4201 ttttggactt tcagtacgtt accggcaaaa tcagtaacgg cttaaagaat cttagctcga
     4261 acgggaagtt ggctgggacc ggggctctta cgggtgaagc atccgagtta atgccaggga
     4321 gccgcacgta ccgtaaagta gacccgccgg acgtagagca gttaggccgt tcatcatgga
     4381 ctttgttaca ttctgtcgcc gcatcgtacc ctgcacagcc taccgatcag caaaagggag
     4441 agatgaagca atttttaaat atcttttctc acatttaccc gtgcaattgg tgcgcgaaag
     4501 actttgaaaa atacattcgt gaaaacgcgc cacaagtcga gtcacgtgag gagttaggtc
     4561 gttggatgtg cgaggcccac aacaaagtca acaaaaagtt acgtaagccg aagttcgact
     4621 gtaatttctg ggaaaagcgc tggaaggatg gctgggacga gtgataatcg accaggcatc
     4681 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     4741 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     4801 tgtttgccct atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     4861 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tatggacgca
     4921 cctgaagagg aagatcatgt tttagtcttg cgtaagtcta acttcgcaga ggccttggct
     4981 gctcataaat accttttggt agagttttac gcaccttggt gcgggcattg caaggctctt
     5041 gctcccgaat atgcgaaagc cgctggaaaa ctgaaggcgg aaggatcgga aatccgcctt
     5101 gccaaggtgg atgcgacgga ggagtcagac ctggcccagc aatacggggt ccgtgggtac
     5161 ccgacaatta aattcttccg caatggtgac acagcatcac cgaaagaata cacggcgggg
     5221 cgtgaggccg atgacatcgt taactggctt aagaaacgca ctgggccagc agcgactacc
     5281 ctgccagatg gggcagcagc tgaaagtttg gtggaatctt ccgaagtggc cgtaattggg
     5341 ttttttaagg acgtagagtc agactcggca aagcaattcc tgcaagctgc tgaggcaatt
     5401 gacgatattc cattcggtat cacctcaaat tcggacgtat tctctaagta tcagctggac
     5461 aaagatggcg tagtgttatt taagaaattt gatgagggtc gtaataactt cgagggcgaa
     5521 gtgaccaaag agaacctttt ggattttatt aagcacaatc aactgccact ggttatcgaa
     5581 tttacagaac agacggcccc taaaatcttc ggcggtgaaa tcaagaccca cattttgctg
     5641 ttcctgccta agagtgtctc ggactacgat ggcaaactga gcaactttaa gaccgcggct
     5701 gagagtttta aggggaaaat ccttttcatt ttcatcgaca gtgatcacac tgataatcaa
     5761 cgcattttgg agtttttcgg tctgaaaaag gaagagtgcc cagcagtgcg tttgattacg
     5821 cttgaagaag aaatgacgaa atacaagcct gagtctgaag aattgaccgc tgagcgcatt
     5881 acagagttct gtcatcgctt cctggagggc aaaatcaaac ctcaccttat gtcgcaggaa
     5941 ttaccagaag attgggataa acaacccgtc aaagtattag tgggtaaaaa tttcgaagat
     6001 gtagcgttcg acgaaaagaa gaacgttttc gtagagttct atgcaccgtg gtgtggtcac
     6061 tgtaagcagc tggcacctat ttgggataag ctgggagaaa cgtataaaga tcacgaaaat
     6121 atcgttatcg cgaagatgga ttcaactgcg aacgaagtgg aagcggtgaa ggttcactct
     6181 tttccaacac ttaagttttt ccctgcttcg gcggaccgta cagtgatcga ctataacggc
     6241 gaacgtactc ttgatgggtt taaaaaattt ttagagagtg gtggccagga tggggcaggt
     6301 gacgacgacg atcttgaaga cttagaagaa gcggaggaac ccgacatgga agaagatgac
     6361 gaccagaaag cagtaaagga tgaattgtga taatcgacca ggcatcaaat aaaacgaaag
     6421 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     6481 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatat
//
