LOCUS       sequence-358137-        3082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HiFi-compatible destination vector for secreted overexpression of 
            thrombin-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 3082)
  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 3082)
  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-CR-PA
FEATURES             Location/Qualifiers
     source          1..3082
                     /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..1142
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     polyA_signal    1583..1712
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(1847..2435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1927..1946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2692..2710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2714..2723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2720..3082,1..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 atctctggtg ccacgcggat
     1141 cggccgccca gcatgacgag gccgtggata acaagttcaa taaggagcag cagaacgcct
     1201 tctacgagat cctgcacctg cccaacctga atgaggagca gcgcaatgcc ttcattcaga
     1261 gcctgaagga cgatccatcc cagtcggcca acctgctggc cgaggccaag aagctgaatg
     1321 acgcccaggc cccgaaggtg gacaacaagt ttaataagga gcagcagaat gccttttatg
     1381 agatcctgca tctgccaaat ctgaacgagg agcagcgcaa cgcctttatc cagtccctga
     1441 aggatgatcc aagccagtcc gccaatttat tagctgaagc caagaagctg aacgatgccc
     1501 aggcccccaa ggtggatgcc aactcctcga gcgtgccata aaccggtcat atgcggccgc
     1561 ggatcgatcg atatctgact aaatcttagt ttgtattgtc atgttttaat acaatatgtt
     1621 atgtttaaat atgtttttaa taaattttat aaaataattt caacttttat tgtaacaaca
     1681 ttgtccattt acacactcct ttcaagcgcg tgggactcga tgctcggcgc cactcaaagg
     1741 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1801 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1861 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1921 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1981 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2041 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2101 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2161 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2221 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2281 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2341 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2401 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttgcatgcg ctaagcgggc
     2521 tttataaaac gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt
     2581 gtggatactc ctcccgacac cgaattaatt cggatctctg caagggattt tggtcatgaa
     2641 caataaaact gtctgcttac ataaacagta atacaagggg tgttcatagt ataatacgac
     2701 tcactatagg agggccacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
     2761 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttcagcc gggacttcgt
     2821 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
     2881 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
     2941 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
     3001 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
     3061 ctgcgtgcac ttcgtggccg ag
//
