LOCUS       Exported                5485 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of human VAP-A fused to GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFPC1-hVAP-A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5485)
  AUTHORS   Alpy F, Rousseau A, Schwab Y, Legueux F, Stoll I, Wendling C, 
            Spiegelhalter C, Kessler P, Mathelin C, Rio MC, Levine TP, Tomasetto
            C
  TITLE     STARD3 or STARD3NL and VAP form a novel molecular tether between 
            late endosomes and the ER.
  JOURNAL   J Cell Sci. 2013 Dec 1;126(Pt 23):5500-12. doi: 10.1242/jcs.139295. 
            Epub 2013 Oct 8.
  PUBMED    24105263
REFERENCE   2  (bases 1 to 5485)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5485
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        47..350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        351..554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     505..525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      593..602
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             599..1315
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             599..1315
                     /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(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(905..924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1252..1273
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2259..2380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2296..2315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2350..2369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2387..2842)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2524..2545)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2736..2755
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2869..2973
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2975..3332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2998..3018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3183..3318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3245..3264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3367..4161
                     /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(3421..3440)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4031..4050
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4349..4368)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4393..4440
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4769..5357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5258..5277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
       61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      541 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
      601 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      721 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      901 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1081 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1201 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     1321 actcagatct cgagctcaag cttcgaattg tgcgtccgcc tcaggggcca tggcgaagca
     1381 cgagcagatc ctggtcctcg atccgcccac agacctcaaa ttcaaaggcc ccttcacaga
     1441 tgtagtcact acaaatctta aattgcgaaa tccatcggat agaaaagtgt gtttcaaagt
     1501 gaagactaca gcacctcgcc ggtactgtgt gaggcccaac agtggaatta ttgacccagg
     1561 gtcaactgtg actgtttcag taatgctaca gccctttgac tatgatccga atgaaaagag
     1621 taaacacaag tttatggtac agacaatttt tgctccacca aacacttcag atatggaagc
     1681 tgtgtggaaa gaggcaaaac ctgatgaatt aatggattcc aaattgagat gcgtatttga
     1741 aatgcccaat gaaaatgata aattgaatga tatggaacct agcaaagctg ttccactgaa
     1801 tgcatctaag caagatggac ctatgccaaa accacacagt gtttcactta atgataccga
     1861 aacaaggaaa ctaatggaag agtgtaaaag acttcaggga gaaatgatga agctatcaga
     1921 agaaaatcgg cacctgagag atgaaggttt aaggctcaga aaggtagcac attcggataa
     1981 acctggatca acctcaactg catccttcag agataatgtc accagtcctc ttccttcact
     2041 tcttgttgta attgcagcca ttttcattgg attctttcta gggaaattca tcttgtagca
     2101 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     2161 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2221 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2341 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2401 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2461 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2521 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2581 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2641 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2701 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2761 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2821 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2941 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3241 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3301 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3361 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3421 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3481 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3541 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3601 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3661 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3721 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3781 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3841 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3901 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3961 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4021 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4081 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4141 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4201 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4261 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4321 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4381 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4441 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4501 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4561 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5461 ccgccatgca ttagttatta atagt
//
