LOCUS       sequence-358138-        3088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HiFi-compatible destination vector for secreted overexpression of 
            3C-cleavable, C-terminal Protein A tagged proteins with ExpreS2 
            Platform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3088)
  AUTHORS   Coker JA, Katis VL, Fairhead M, Schwenzer A, Clemmensen SB, Frandsen
            BU, de Jongh WA, Gileadi O, Burgess-Brown NA, Marsden BD, Midwood 
            KS, Yue WW
  TITLE     FAS2FURIOUS: Moderate-Throughput Secreted Expression of Difficult 
            Recombinant Proteins in Drosophila S2 Cells.
  JOURNAL   Front Bioeng Biotechnol. 2022 May 5;10:871933. doi: 
            10.3389/fbioe.2022.871933. eCollection 2022.
  PUBMED    35600892
REFERENCE   2  (bases 1 to 3088)
  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: "Front 
            Bioeng Biotechnol."; date: "2022-05-5"; volume: "10:871933. doi"; 
            pages: " 10.3389/fbioe.2022.871933. eCollection 2022"
COMMENT     Sequence Label: pExpreS2-1-C3-PA
FEATURES             Location/Qualifiers
     source          1..3088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    76..197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(113..132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     intron          910..1042
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     sig_peptide     1065..1118
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1125..1148
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    1589..1718
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(1853..2441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1933..1952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2698..2716
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2720..2729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2726..3088,1..12)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 gagcaggact gaccgacgcc gaccaacacc gccggtccga cggcggccca cgggtcccag
       61 gggggtcgac ctcgaaactt gtttattgca gcttataatg gttacaaata aagcaatagc
      121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
      181 ctcatcaatg tatcttatca tgtcttcacg taataagtgt gcggctagca gtcaactagt
      241 ccaaactcat caatgtatct tatcatgtct tcacgtaata agtgtgcggc tagcagtcaa
      301 ctactagtga atgccctact agaagatgtg tgttgcacaa aatgtccctg gaataaccaa
      361 tttgaagtgc agatagcagt aaacgtaagc taatatgaat attatttaac tgtaatgttt
      421 taatatcgct ggacattact aataaaccca ctataaacac atgtacatat gtatgttttg
      481 gcatacaatg agtagttggg gaaaaaatgt gtaaaagcac cgtgaccatc acagcataaa
      541 gataaccagc tgaagtatcg aatatgagta acccccaaat tgaatcacat gccgcaactg
      601 ataggaccca tggaagtaca ctcttcatgg cgatatacaa gacacacaca agcacgaaca
      661 cccagttgcg gaggaaattc tccgtaaatg aaaacccaat cggcgaacaa ttcataccca
      721 tatatggtaa aagttttgaa cgcgagcgaa cttaagagcg ccggagtata aatagaggcg
      781 cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa
      841 agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagaa gcttggtacc
      901 ctcgagctcg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
      961 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     1021 cactttgcct ttctctccac aggaattctg gatccgatct caatatgaag ttatgcatat
     1081 tactggccgt cgtggccttt gttggcctct cgctcgggag atctttggag gttctgtttc
     1141 agggtccagc cgcccagcat gacgaggccg tggataacaa gttcaataag gagcagcaga
     1201 acgccttcta cgagatcctg cacctgccca acctgaatga ggagcagcgc aatgccttca
     1261 ttcagagcct gaaggacgat ccatcccagt cggccaacct gctggccgag gccaagaagc
     1321 tgaatgacgc ccaggccccg aaggtggaca acaagtttaa taaggagcag cagaatgcct
     1381 tttatgagat cctgcatctg ccaaatctga acgaggagca gcgcaacgcc tttatccagt
     1441 ccctgaagga tgatccaagc cagtccgcca atttattagc tgaagccaag aagctgaacg
     1501 atgcccaggc ccccaaggtg gatgccaact cctcgagcgt gccataaacc ggtcatatgc
     1561 ggccgcggat cgatcgatat ctgactaaat cttagtttgt attgtcatgt tttaatacaa
     1621 tatgttatgt ttaaatatgt ttttaataaa ttttataaaa taatttcaac ttttattgta
     1681 acaacattgt ccatttacac actcctttca agcgcgtggg actcgatgct cggcgccact
     1741 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1801 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2041 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2281 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg catgcgctaa
     2521 gcgggcttta taaaacgggc tgcgggacca gttttcatat cactaccgtt tgagttcttg
     2581 tgctgtgtgg atactcctcc cgacaccgaa ttaattcgga tctctgcaag ggattttggt
     2641 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt catagtataa
     2701 tacgactcac tataggaggg ccaccatggc caagttgacc agtgccgttc cggtgctcac
     2761 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tcagccggga
     2821 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     2881 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     2941 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     3001 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     3061 cggcaactgc gtgcacttcg tggccgag
//
