LOCUS       sequence-424762-        5086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5086)
  AUTHORS   Paumet F, Le Mao J, Martin S, Galli T, David B, Blank U, Roa M
  TITLE     Soluble NSF attachment protein receptors (SNAREs) in RBL-2H3 mast 
            cells: functional role of syntaxin 4 in exocytosis and 
            identification of a vesicle-associated membrane protein 8-containing
            secretory compartment.
  JOURNAL   J Immunol. 2000 Jun 1;164(11):5850-7.
  PUBMED    10820264
REFERENCE   2  (bases 1 to 5086)
  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: "J 
            Immunol."; date: "2000-06-1"; volume: "164(11)"; pages: "5850-7"
COMMENT     Sequence Label: pEGFP VAMP8
FEATURES             Location/Qualifiers
     source          1..5086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             44..838
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             44..838
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(98..117)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     708..727
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1026..1045)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1070..1117
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1446..2034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1935..1954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2209..2512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2513..2716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2667..2687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2755..2764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2761..2763,2764..2766,2767..3477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2761..2763,2764..2766,2767..3477)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2806..2827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3067..3086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3414..3435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4022..4143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4059..4078)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4113..4132
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4150..4605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4287..4308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4499..4518
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4632..4736
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4738..5086,1..9)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4738..4929
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4761..4781)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4946..5081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
       61 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      121 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
      181 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
      241 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
      301 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
      361 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
      421 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
      481 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
      541 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
      601 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
      661 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
      721 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
      781 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
      841 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
      901 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
      961 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     1021 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     1081 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     1141 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     1201 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     1261 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     1321 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1381 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2101 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     2161 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2221 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2281 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2341 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2401 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2461 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     2521 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     2581 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2641 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2701 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
     2761 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2821 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2881 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2941 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3001 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3061 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3121 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3181 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3241 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3301 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3361 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3421 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtac
     3481 tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgagctc ggatccacta
     3541 gtaacggccg ccagtgtgct ggaattcgcc cttatggagg aagccagtga aggtggagga
     3601 aatgatcgtg tgcggaacct gcaaagtgag gtggagggag ttaagaatat tatgacccag
     3661 aatgtggagc ggatcctggc ccggggggaa aacttggaac atctccgcaa caagacagag
     3721 gatctggaag ccacatctga gcacttcaag acgacatcgc agaaggtggc tcgaaaattc
     3781 tggtggaaga acgtgaagat gattgtcctt atctgcgtga ttgtttttat catcatcctc
     3841 ttcattgtgc tctttgccac tggtgccttc tcaagggcga attctgcaga tgggatccac
     3901 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3961 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4021 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4081 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4141 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4201 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4261 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4321 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4381 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4441 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4501 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4561 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4621 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4681 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4741 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4921 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4981 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5041 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggc
//
