LOCUS       sequence-360135-        3214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HiFi-compatible destination vector for secreted overexpression of 
            3C-cleavable, C-terminal Protein A-10His-TwinStrep tagged proteins 
            with ExpreS2 Platform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3214)
  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 3214)
  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-10H-SIII
FEATURES             Location/Qualifiers
     source          1..3214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          630..762
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     sig_peptide     785..838
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             845..868
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1268..1297
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(1307..1330,1331..1366,1367..1390)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    1435..1564
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(1699..2287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1779..1798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2544..2562
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2566..2575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2572..2946
                     /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"
     polyA_signal    3010..3131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3047..3066)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 agtgtgcggc tagcagtcaa ctactagtga atgccctact agaagatgtg tgttgcacaa
       61 aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc taatatgaat
      121 attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca ctataaacac
      181 atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt gtaaaagcac
      241 cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta acccccaaat
      301 tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg cgatatacaa
      361 gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg aaaacccaat
      421 cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgagcgaa cttaagagcg
      481 ccggagtata aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag
      541 tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca
      601 atctgcagaa gcttggtacc ctcgagctcg taagtatcaa ggttacaaga caggtttaag
      661 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
      721 ctattggtct tactgacatc cactttgcct ttctctccac aggaattctg gatccgatct
      781 caatatgaag ttatgcatat tactggccgt cgtggccttt gttggcctct cgctcgggag
      841 atctttggag gttctgtttc agggtccagc cgcccagcat gacgaggccg tggataacaa
      901 gttcaataag gagcagcaga acgccttcta cgagatcctg cacctgccca acctgaatga
      961 ggagcagcgc aatgccttca ttcagagcct gaaggacgat ccatcccagt cggccaacct
     1021 gctggccgag gccaagaagc tgaatgacgc ccaggccccg aaggtggaca acaagtttaa
     1081 taaggagcag cagaatgcct tttatgagat cctgcatctg ccaaatctga acgaggagca
     1141 gcgcaacgcc tttatccagt ccctgaagga tgatccaagc cagtccgcca atttattagc
     1201 tgaagccaag aagctgaacg atgcccaggc ccccaaggtg gatgccaact cctcgagcgt
     1261 gccatcgcac catcatcacc atcaccatca ccaccatggt tctggttgga gtcacccaca
     1321 attcgagaaa ggtggtggct ctggtggcgg ttcaggcggc tccgcctggt cccatccaca
     1381 gtttgaaaag tgaaccggtc atatgcggcc gcggatcgat cgatatctga ctaaatctta
     1441 gtttgtattg tcatgtttta atacaatatg ttatgtttaa atatgttttt aataaatttt
     1501 ataaaataat ttcaactttt attgtaacaa cattgtccat ttacacactc ctttcaagcg
     1561 cgtgggactc gatgctcggc gccactcaaa ggcggtaata cggttatcca cagaatcagg
     1621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1861 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1921 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2101 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2341 acgttaaggg attttgcatg cgctaagcgg gctttataaa acgggctgcg ggaccagttt
     2401 tcatatcact accgtttgag ttcttgtgct gtgtggatac tcctcccgac accgaattaa
     2461 ttcggatctc tgcaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag
     2521 taatacaagg ggtgttcata gtataatacg actcactata ggagggccac catggccaag
     2581 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     2641 accgaccggc tcgggttcag ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     2701 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     2761 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     2821 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     2881 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     2941 gactgaccga cgccgaccaa caccgccggt ccgacggcgg cccacgggtc ccaggggggt
     3001 cgacctcgaa acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3061 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3121 aatgtatctt atcatgtctt cacgtaataa gtgtgcggct agcagtcaac tagtccaaac
     3181 tcatcaatgt atcttatcat gtcttcacgt aata
//
