LOCUS       sequence-449610-        5130 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 5130)
  AUTHORS   Mallard F, Tang BL, Galli T, Tenza D, Saint-Pol A, Yue X, Antony C, 
            Hong W, Goud B, Johannes L
  TITLE     Early/recycling endosomes-to-TGN transport involves two SNARE 
            complexes and a Rab6 isoform.
  JOURNAL   J Cell Biol. 2002 Feb 18;156(4):653-64. Epub 2002 Feb 11.
  PUBMED    11839770
REFERENCE   2  (bases 1 to 5130)
  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 Cell 
            Biol."; date: "2002-02-18"; volume: "156(4)"; pages: "653-64. Epub 
            2002 Feb 11"
COMMENT     Sequence Label: pEGFP VAMP4
FEATURES             Location/Qualifiers
     source          1..5130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        158..349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     306..326
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(351..455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      482..937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(569..588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     779..800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    944..1065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(955..974)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1009..1028
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1654..2364,2365..2367,2368..2370))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1654..2364,2365..2367,2368..2370))
                     /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(1696..1717)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2045..2064
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2304..2325
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2367..2376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2415..2618)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2444..2464)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2619..2922
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3097..3685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3177..3196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4014..4061
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4086..4105
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4293..5087)
                     /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             complement(4293..5087)
                     /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(4404..4423)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5014..5033
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5122..5130,1..349))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
       61 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      241 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      301 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
      361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
      421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
      481 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      541 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      601 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      661 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      781 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      841 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      901 aacgcgaatt ttaacaaaat attaacgctt acaatttacg cgttaagata cattgatgag
      961 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1021 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1081 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1141 ctctacaaat gtggtatggc tgattatgat cagttatcta gatccggtgg atcctcaagt
     1201 acggtatttc atgactataa gaatgataat cactagcaaa aggatagcag caaccaaagc
     1261 catgatggct tttattttgc atccacgcca ccacatttgc cttcgaagtt gtttggatct
     1321 gttgctaaaa gctgttgcat tatccgataa gctttctgat ttgtcctgta gttcatctag
     1381 tctctcccct ctctcaatta cctttgtaat attttcttgc atgacatcaa taacttcatc
     1441 cacttgattc tgaacatgct taattttatc atttctaggt ccaaatcttg gtccagatgg
     1501 tccccttaga aaaaagtcct cttcttcatc tgaatcatct tccaaaagat ttctcctttc
     1561 acttttcaca gaacctgtga catcatcatc attgaggtgg cgcttaaact tgggaggcat
     1621 cagaattcga agcttgagct cgagatctga gtacttgtac agctcgtcca tgccgagagt
     1681 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1741 tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1801 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1861 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1921 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1981 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     2041 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     2101 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     2161 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     2221 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     2281 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     2341 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcggatctga
     2401 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     2461 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     2521 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     2581 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     2641 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     2701 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     2761 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     2821 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     2881 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     2941 ctatgaacta atgaccccgt aattgattac tattaataac taatgcatgg cggtaatacg
     3001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3841 acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc
     3901 cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg
     3961 tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa
     4021 cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
     4081 cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag
     4141 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     4201 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     4261 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     4321 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4381 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4441 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4501 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
     4561 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4621 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4681 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4741 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4801 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4861 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     4921 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4981 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     5041 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     5101 catcctgtct cttgatcgat ctttgcaaaa
//
