LOCUS       sequence-349902-        6491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression plasmid expressing PDI and Erv2 from Fusarium oxysporum
            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 6491)
  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 6491)
  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-T7FunCyDisCo
FEATURES             Location/Qualifiers
     source          1..6491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1741..1760
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1788..1806
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1807..1831
                     /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             1846..1868
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2441..2512
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2528..2555
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2584..2602
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2603..2627
                     /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             2642..2664
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      4152..4223
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4239..4266
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        complement(join(4417..4422,4423..4439,4440..4445))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4557..5102
                     /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     5003..5022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5219..5236
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5617..5719
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5720..6379
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5786..5805)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac
       61 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
      121 ccccgtggag gtaataattg acgatatgat catttattct gcctcccaga gcctgataaa
      181 aacggttagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
      241 gcgatgcaga tccggaacat aatggtgcag ggcgcttgtt tcggcgtggg tatggtggca
      301 ggccccgtgg ccgggggact gttgggcgct gccggcacct gtcctacgag ttgcatgata
      361 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagcta
      421 ccggcagcgg tgcggactgt tgtaactcag aataagaaat gaggccgctc atggcgttga
      481 ctctcagtca tagtatcgtg gtatcaccgg ttggttccac tctctgttgc gggcaacttc
      541 agcagcacgt aggggacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
      601 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
      661 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
      721 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc
      781 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg
      841 cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag
      901 cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa
      961 cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt
     1021 gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct
     1081 ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
     1141 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
     1201 gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc
     1261 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     1321 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     1381 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     1441 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     1501 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgacgct ctcccttatg
     1561 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     1621 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     1681 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     1741 ggtgatgtcg gcgatatagg catctctata tctctatccc gcgaaattaa tacgactcac
     1801 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
     1861 gaaggagata taccatatga gcggcccgag tcgttcaatc cttcccaaat accagaacga
     1921 tgtagagtta ccgcttaagg aagccccacg ctcggagttc gccgctgacc ttggtgctct
     1981 tccggccggt ttgttggatg ggaactcgat tgctccaaaa ttggagaacg caacacttaa
     2041 ggcggagttg gggcatgcga cttggaaatt cttacacacg atgatggccc gcttccctga
     2101 caaaccgacc aaagatgacc gcatggcact tgaaaccttt atgcacttat tcgcccgctt
     2161 atatccttgc ggtcaatgcg cggctcactt tcaaaagtta ctggcacaat accctccaca
     2221 aacatcatcc cgcaatgccg cggccggatg gttgtgcttt gctcataata tcgtcaatga
     2281 acgcgtccac aaaccgttat tcgattgcga gaatatcggg gacttctacg attgcgggtg
     2341 cggcgacaaa gacaagaagg agggtggagc agaaaccact gcagccgcgc aagccgaagg
     2401 cccagaccag gagcttcata aacactaatc gaccaggcat caaataaaac gaaaggctca
     2461 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     2521 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atgtttgccc tatcccgcga
     2581 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     2641 ttttgtttaa ctttaagaag gagatatacc atatggcaga tagtgacgtc caccaactga
     2701 ctaaggacac gtttgaggag tttgttaaaa gtaacgactt ggtacttgct gaattttttg
     2761 ctccgtggtg tggtcattgt aaagcgttgg cacccgaata tgaggaagcc gctaccaccc
     2821 tgaaggagaa gaatatcaag ctggcaaaga tcgactgtac cgaagaatcc gacctgtgta
     2881 aggatcaggg ggtcgagggt tatcccactt taaaagtgtt tcgcggcttg gagaacgtca
     2941 cgccatactc ggggcaacgc aaggcggcgg gtatcacatc atatatgatt aagcagtcct
     3001 tacctgcggt atcaattttg acgaaggata cacttgagga atttaaaact gcggacaagg
     3061 ttgttgttgt agcgtactta aatgcagacg acaagtctag caacgaaact ttctctaagt
     3121 tagctgaagg gttacgtgac acgtatctgt tcggtggggt caatgatgct gccgtcgcag
     3181 aagctgaggg cgttaaggcg ccagctctgg ttgtctataa atcgtttgat gaaggtaaga
     3241 acacattcac tgagaagttc gaagaagatg cgattgccag ctttatcacc accagtgcaa
     3301 cgcccttgat cggggaagtg ggtcctgaaa catacgctgg gtacatgtcg gcgggcatcc
     3361 ccctggccta catcttctca gaaacaccag aagagcgtaa ggaactgggt gacgcattga
     3421 aacctatcgc ggaaaagttt aaaggtaaaa ttaatttcgc aactattgac gcgaaggctt
     3481 tcggggcaca cgcggggaat ttaaacttga aggccgacaa gtttccttcg tttgcgattc
     3541 aagaagtagt caaaaaccag aaatttccgt ttgatcaaga gaaggagatc acccatgaca
     3601 acattgctaa gttcgtggaa gattttgcgg ctggaaagat cgaaccctcc attaagtctg
     3661 agcctattcc tgaaacgcag gagggaccag tcacggtagt cgtggccaaa tcgtacaacg
     3721 acatcgtact tgatgacacg aaagacgtgt tgatcgaatt ctacgctccc tggtgcggcc
     3781 actgtaaggc attggcaccc aaatacgagg acttggcgtc acaatttgcc gcgtccgagt
     3841 tcaaagataa ggtagtcatc gccaaggtgg acgcaaccct taacgatgta cctgatgaga
     3901 tccagggatt ccctacaatc aagttgtatg cagcaggagc caaagacgca ccggtcacat
     3961 accaaggttc acgcaccgtc gaagatttgg ccaatttcat taaagagaac ggcaaataca
     4021 aggctgagtt gcctgtaaag gaggagggga ctgaggaggc cgcgcccgct gcgagtgaag
     4081 aaaagaagga ggaaaagaag gatgcggaag aagaagacgt ccatgacgaa ttgtgataat
     4141 cgaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4201 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     4261 tctgcgttta tatgttccgg ctagctgcag tcataagtta ggtacctagc tgcagtcata
     4321 agggacttag ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     4381 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     4441 gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     4501 gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga
     4561 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     4621 ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga
     4681 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     4741 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     4801 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     4861 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     4921 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     4981 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5041 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     5101 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     5161 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     5221 cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg
     5281 tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca
     5341 tagtaagcca gtatacactc cgctagcgct gatgtccggc ggtgcttttg ccgttacgca
     5401 ccaccccgtc agtagctgaa caggagggac agctgataga aacagaagcc actggagcac
     5461 ctcaaaaaca ccatcataca ctaaatcagt aagttggcag catcacccga cgcactttgc
     5521 gccgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct
     5581 gttgataccg ggaagccctg ggccaaaaaa agacgttgat cggcacgtaa gaggttccaa
     5641 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     5701 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     5761 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     5821 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     5881 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     5941 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     6001 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     6061 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     6121 ccctaaaggg tttattgaga atatgttttt cgtgtcagcc aatccctggg tgagtttcac
     6181 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     6241 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     6301 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     6361 gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg
     6421 ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac
     6481 ccggtcgtcg g
//
