LOCUS       sequence-206171-        6174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SNAP47 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6174)
  AUTHORS   Kuster A, Nola S, Dingli F, Vacca B, Gauchy C, Beaujouan JC, Nunez 
            M, Moncion T, Loew D, Formstecher E, Galli T, Proux-Gillardeaux V
  TITLE     The Q-soluble N-Ethylmaleimide-sensitive Factor Attachment Protein 
            Receptor (Q-SNARE) SNAP-47 Regulates Trafficking of Selected 
            Vesicle-associated Membrane Proteins (VAMPs).
  JOURNAL   J Biol Chem. 2015 Nov 20;290(47):28056-69. doi: 
            10.1074/jbc.M115.666362. Epub 2015 Sep 10.
  PUBMED    26359495
REFERENCE   2  (bases 1 to 6174)
  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"; doi: 
            "10.1074/jbc.M115.666362"; journalName: "J Biol Chem"; date: 
            "2015-11-20- 20"; volume: "290"; issue: "47"; pages: "28056-69"
COMMENT     Sequence Label: pEGFP SNAP47
FEATURES             Location/Qualifiers
     source          1..6174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2902..3023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2939..2958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2993..3012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3030..3485)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3167..3188)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3379..3398
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3512..3616
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3618..3975
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3618..3809
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3641..3661)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3826..3961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4010..4804
                     /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             4010..4804
                     /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(4064..4083)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4674..4693
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4992..5011)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5036..5083
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5412..6000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5901..5920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt actcagatct cgagctcaag cttcgaattc gtcgactgga tccggtaccg
     1321 aggagatctg ccgccgcgat cgccatgcgc gcggctcgcc gcggtcttca ctgcgcaggc
     1381 gccgagcggc cgaggcgccg cggtcggctc tgggactcgt ctggcgtccc tcagaggcag
     1441 aagaggcctg gaccttggcg cacacagacc caggaacaga tgagcaggga tgtctgcatc
     1501 cacacctggc cgtgcaccta ctacctggag cccaagaggc gatgggttac tggacagctg
     1561 tccttaacat cgctgtcgct caggttcatg actgacagca ctggagagat tctggtcagc
     1621 ttccccctct ccagcatagt tgagatcaag aaggaggctt cacattttat cttcagctcc
     1681 atcaccatcc tggagaaggg ccatgccaag cactggttca gctccctgcg gccaagtcga
     1741 aatgtggtct tcagcatcat cgagcatttc tggagggagc tgctgctgtc tcagcctgga
     1801 gccgtggcag acgcatctgt cccaaggacc cggggcgagg agctgacggg actcatggct
     1861 ggatcccaga aacgcctgga ggacacggcg agggtcctgc accaccaggg ccagcagctg
     1921 gacagcgtca tgagaggcct ggacaagatg gagtcagacc tggaggtggc ggacagattg
     1981 ctgacagaac tggaatctcc tgcttggtgg ccctttagct ccaagctttg gaagacacca
     2041 ccggaaacaa agcccaggga agatgtctcc atgaccagtt gtgaaccctt tgggaaagaa
     2101 gggatactga taaaaattcc tgctgttatt tcccacagaa cagagtctca cgttaaacca
     2161 gggaggctca ccgtccttgt gtctgggttg gaaatacatg actccagttc tttgctcatg
     2221 cacaggtttg aaagagaaga cgtggacgac atcaaggtcc actcacctta cgaaattagc
     2281 atccgccagc ggtttattgg aaagccagac atggcctatc gtttgatatc tgccaagatg
     2341 ccagaggtta tccccatttt agaagtgcag ttcagcaaga agatggagct gttagaagat
     2401 gcattggtgc tcagaagcgc aagaacctct tcccccgcag agaagagctg ctcagtctgg
     2461 catgcagcat ctgggctgat gggctgtacc ctgcaccgtg agccacccgc aggagaccag
     2521 gagggcacag cactgcacct gcagacaagc ctgccagccc tttctgaggc agatacccag
     2581 gaactaaccc agatcctgag gaggatgaag gggctggccc tggaggccga gagtgagctg
     2641 gagagacaag acgaagccct ggatggcgtt gcagcagctg tggacagggc aaccttgacc
     2701 atcgacaagc acaacaggcg gatgaagagg ctgaccacgc gtacgcggcc gctcgacggt
     2761 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     2821 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2881 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2941 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3001 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3061 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3121 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3181 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3241 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3301 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3361 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3421 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3481 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3541 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3601 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3781 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3841 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3901 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3961 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4021 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4081 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4141 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4201 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4261 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4321 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4381 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4441 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4501 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4561 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4621 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4681 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4741 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4801 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4981 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5041 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5101 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5161 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5221 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5281 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5341 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5401 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5461 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5521 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5581 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5641 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5701 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5761 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5821 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5881 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5941 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6001 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6061 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     6121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
