LOCUS       sequence-457777-        5456 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 5456)
  AUTHORS   Kaiser SE, Brickner JH, Reilein AR, Fenn TD, Walter P, Brunger AT
  TITLE     Structural basis of FFAT motif-mediated ER targeting.
  JOURNAL   Structure. 2005 Jul . 13(7):1035-45.
  PUBMED    16004875
REFERENCE   2  (bases 1 to 5456)
  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: "Structure. 
            2005 Jul . 13(7):1035-45."
COMMENT     Sequence Label: pEGFP-N1-VAPA(1-242)
FEATURES             Location/Qualifiers
     source          1..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      75..530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(162..181)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     372..393
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    537..658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(548..567)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     602..621
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(781..1494,1495..1497,1498..1500))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(784..1494,1495..1497,1498..1500))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(826..847)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1175..1194
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1434..1455
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1497..1506
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2334..2537)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2363..2383)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2538..2841
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3016..3604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3096..3115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3933..3980
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4005..4024
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4212..5006)
                     /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(4212..5006)
                     /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(4323..4342)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4933..4952
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5041..5398)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5055..5190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5207..5398
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5355..5375
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(join(5400..5456,1..48))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
       61 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
      121 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
      181 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
      241 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
      301 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
      361 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
      421 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
      481 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt acgccttaag
      541 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      601 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      661 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      721 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgct
      781 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
      841 gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta
      901 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
      961 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc
     1021 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
     1081 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1141 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1201 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1261 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1321 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1381 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1441 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1501 ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc gaagcttcga
     1561 gatgaatttc cctagaaaga atccaatgaa aatggctgca attacaacca gaagagaagg
     1621 aagaggactg gtgacattat ctctgaagga cacggctgag gtggatccag gtttatccga
     1681 atgtgctacc tttctgagcc gtaggccttc atctctcagg tgtcggtttt cttctgagag
     1741 cttcatcatt tctccctgaa gtcgcttgca ctcttccatc agtttccttg tctccgtgtc
     1801 attgagtgaa acactatgtg gttttggcag gggtccatct tgcttggatg cattcagtgg
     1861 gacagctttg ctaggttcca tgtcgttcag cttatcgttt tcattcggca tctcaaatac
     1921 acatctcagt ttagaatcca ttagttcgtc aggttttgcc tctttccaca cagcttccat
     1981 atctgaaatg tttggtggag caaaaattgt ctgtaccatg aacttatgtt tactcttttc
     2041 gttcggatca tagtcaaagg gctgcagcat tactgagaca gtcacgattg accctgggtc
     2101 aataacccca ctgttgggcc gcacgcagta ccggcggggc gctgtggtct tcactttgaa
     2161 acacactttt ctgtccgatg gattctgcaa tttaagattt gtagtgacta catctgtgaa
     2221 ggggcctttg aatttgaggt ctgaaggagg gtcgaggacc aggatttgct cgtgtttcgc
     2281 catctcgaga tctgagtccg gtagcgctag cggatctgac ggttcactaa accagctctg
     2341 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     2401 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     2461 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     2521 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     2581 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     2641 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     2701 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     2761 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     2821 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     2881 attgattact attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga
     2941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     3781 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     3841 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     3901 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     3961 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     4021 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     4081 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     4141 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     4201 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     4261 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     4321 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     4381 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     4441 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     4501 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     4561 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     4621 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     4681 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     4741 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     4801 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     4861 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     4921 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     4981 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     5041 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     5101 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     5161 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     5221 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5281 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5341 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     5401 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcgg
//
