LOCUS       sequence-358141-        3317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZeoR overexpression of cytosolic GFP in Drosophila ExpreS2ion cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3317)
  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 3317)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..3317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          444..576
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      590..599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(629..657)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1252..1272
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1352..1481
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(1616..2204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1696..1715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2461..2479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2483..2492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2489..2863
                     /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    2927..3048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2964..2983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 atatgtatgt tttggcatac aatgagtagt tggggaaaaa atgtgtaaaa gcaccgtgac
       61 catcacagca taaagataac cagctgaagt atcgaatatg agtaaccccc aaattgaatc
      121 acatgccgca actgatagga cccatggaag tacactcttc atggcgatat acaagacaca
      181 cacaagcacg aacacccagt tgcggaggaa attctccgta aatgaaaacc caatcggcga
      241 acaattcata cccatatatg gtaaaagttt tgaacgcgag cgaacttaag agcgccggag
      301 tataaataga ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca
      361 cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatctgc
      421 agaagcttgg taccctcgag ctcgtaagta tcaaggttac aagacaggtt taaggagacc
      481 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
      541 gtcttactga catccacttt gcctttctct ccacaggaat tctggatccg ccaccatggc
      601 tagcaaagga gaagaactct tcactggagt tgtcccaatt cttgttgaat tagatggtga
      661 tgttaacggc cacaagttct ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      721 acttaccctg aagttcatct gcactactgg caaactgcct gttccatggc caacactagt
      781 cactactctg tgctatggtg ttcaatgctt ttcaagatac ccggatcata tgaaacggca
      841 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaaggacca tcttcttcaa
      901 agatgacggc aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
      961 tagaatcgag ttaaaaggta ttgacttcaa ggaagatggc aacattctgg gacacaaatt
     1021 ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat
     1081 caaagtgaac ttcaagaccc gccacaacat tgaagatgga agcgttcaac tagcagacca
     1141 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     1201 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     1261 tgagtttgta acagctgctg ggattacaca tggcatggat gaactgtaca acatcgatta
     1321 agcggccgcg gatcgatcga tatctgacta aatcttagtt tgtattgtca tgttttaata
     1381 caatatgtta tgtttaaata tgtttttaat aaattttata aaataatttc aacttttatt
     1441 gtaacaacat tgtccattta cacactcctt tcaagcgcgt gggactcgat gctcggcgcc
     1501 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1561 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1621 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1681 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1741 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1801 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1861 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1921 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1981 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2041 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2101 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2161 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2221 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttgcatgcgc
     2281 taagcgggct ttataaaacg ggctgcggga ccagttttca tatcactacc gtttgagttc
     2341 ttgtgctgtg tggatactcc tcccgacacc gaattaattc ggatctctgc aagggatttt
     2401 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttcatagta
     2461 taatacgact cactatagga gggccaccat ggccaagttg accagtgccg ttccggtgct
     2521 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttcagccg
     2581 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     2641 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     2701 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     2761 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     2821 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgaccgacgc cgaccaacac
     2881 cgccggtccg acggcggccc acgggtccca ggggggtcga cctcgaaact tgtttattgc
     2941 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3001 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtcttcac
     3061 gtaataagtg tgcggctagc agtcaactag tccaaactca tcaatgtatc ttatcatgtc
     3121 ttcacgtaat aagtgtgcgg ctagcagtca actactagtg aatgccctac tagaagatgt
     3181 gtgttgcaca aaatgtccct ggaataacca atttgaagtg cagatagcag taaacgtaag
     3241 ctaatatgaa tattatttaa ctgtaatgtt ttaatatcgc tggacattac taataaaccc
     3301 actataaaca catgtac
//
