LOCUS       sequence-348379-        7606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VPS50-GFP is expressed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7606)
  AUTHORS   Schindler C, Chen Y, Pu J, Guo X, Bonifacino JS
  TITLE     EARP is a multisubunit tethering complex involved in endocytic 
            recycling.
  JOURNAL   Nat Cell Biol. 2015 May;17(5):639-50. doi: 10.1038/ncb3129. Epub 
            2015 Mar 23.
  PUBMED    25799061
REFERENCE   2  (bases 1 to 7606)
  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: "10.1038/ncb3129"; 
            journalName: "Nat Cell Biol"; date: "2015-05"; volume: "17"; issue: 
            "5"; pages: "639-50"
COMMENT     Sequence Label: pEGFP-N1 VPS50
FEATURES             Location/Qualifiers
     source          1..7606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..328
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        329..532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     483..503
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3510..3519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3516..3518,3519..3521,3522..4235)
                     /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             join(3516..3518,3519..3521,3522..4232)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3561..3582)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3822..3841)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4169..4190
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4358..4479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4395..4414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4449..4468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4486..4941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4623..4644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4835..4854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4968..5072
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5074..5431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5074..5265
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5097..5117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5282..5417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5466..6260
                     /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             5466..6260
                     /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(5520..5539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6130..6149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6448..6467)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6492..6539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6868..7456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7357..7376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
       61 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      121 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      181 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      541 gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct tcgaattctg
      601 cagtcgacat gcaaaaaatc aaatctctca tgacccggca gggtctgaag agccctccgg
      661 agagcctcaa tgatcttggt gcctttgaga gccttcgggt ccctggaaag gaggaattca
      721 gggagcttcg agaacagcca agcgaccctc aagctgagca agagttgatt aatagcattg
      781 aacaagtcta tttttctgcg gatccatttg atattgttaa atatgagctg gagaagcttc
      841 ctcctgttct caatctgcaa gagttagagg agtacagaga caaattaaag caacaacaat
      901 ctgcagtctc taaaaaagtg gcagatttga ttcttgaaaa gcagcctgct tatgtaaagg
      961 aacttgaaag agtcacatca ctgcagacag gtcttcagtt ggctgctgtc atctgcacaa
     1021 atgggaggag gcacttgaat attgcaaagg aaggttttac tcaggctagc ttaggccttc
     1081 ttgcaaatca aaggaagcgt cagttgctaa ttggacttct gaagtcgctg agaaccataa
     1141 aaacattgca aagaactgac atacgcttaa gtgaaatgct ggaggaagag gactatccag
     1201 gagctattca gctgtgccta gaatgtcaga aggctgccag cacttttaaa cactacagct
     1261 gtataagtga gctgaattca aagctgcaag atactttgga gcaaattgag gaacagttgg
     1321 atgtagctct ttccaaaatc tgcaagaatt ttgacattaa ccattatacc aaggttcagc
     1381 aagcttaccg gcttcttgga aaaacacaga ctgcgatgga tcagcttcat atgcacttca
     1441 ctcaagccat tcacaacacc gtatttcagg ttgtgcttgg ctatgtggag ctgtgtgcag
     1501 gaaacacgga tacaaaattc cagaagcttc agtataaaga tctctgtacg catgtgacac
     1561 cagacagcta tattccatgc ctggcagacc tgtgcaaagc actgtgggaa gtcatgctca
     1621 gctactacag aaccatggag tggcacgaga aacatgacaa tgaggaaaca gctgcagctg
     1681 ctgaaggaag caatgtgatg agtactgagg aggccacttt tgaccgtggc tatgtaaaga
     1741 agaagttaga gcacggcctt acccggatat ggcaggatgt tcaactaaaa gtaaaaacct
     1801 acttgcttgg aactgattta tctatattca aatatgatga tttcatcttt gttctggata
     1861 ttgtcagcag gttgatgcaa gttggagaag aattttgtgg tagcaagtct gaagttttgc
     1921 aggagtccat tagaaaacaa agtgtcaatt acttcaagaa ccaccatagg atacgacttg
     1981 atgaactgag aatgttctta gagaatgaga catgggagct ttgccctgtt aagtcaaatt
     2041 ttagcatctt gcagcttcat gagtttaaat tcctggaaca gtctcgctcc ccatcagttt
     2101 cacctagtaa gcagccatca gcaacttctt caaagcctgt gactttgttt gagcagtact
     2161 gcagtggtgg gaatccattt gaaatacaag ccgaccacaa agatgaagaa actgaagatg
     2221 tcttagcctc taatgggtat gagtctgatg agcaagagaa gagcgcctat caggactatg
     2281 acagtgacag cgacgtccct gaggagctga agcgggatta cgtggacgag cagaccggcg
     2341 acgtgcccgt gaagagtgtt tcccgagaga ccctgaagag caggaagaag tcggattaca
     2401 gtctaaacaa agtgaatgca cccatcttaa caaacacaac actgaatgta ataaggcttg
     2461 ttggaaagta catgcagatg atgaacatcc ttaagccaat cgccttcgat gtgatccatt
     2521 tcatgtctca gctctttgac tattacttgt atgccatata tacctttttt ggtcggaatg
     2581 attcattgga atcaacagga cttggcctta gtagtagcag actaaaaaca actctaaaca
     2641 gaattcaaga aagccttatt gacctggagg gctcagctga tccaactgcc actctcacag
     2701 cagccgagga aagaaaggag aaggtaccca gcccacacct caatcagcta gtgattttga
     2761 catctgggga tacattgtat gggttggcag agagagtggt agccacagaa tccttagtgt
     2821 tcttggctga gcagtttgag ttccttcagc cacatttgga tgctgtgatg cctgcagtca
     2881 aaaagccctt cctgcagcag ttttactccc agacagtctc aactgccagt gaactgcgca
     2941 agcccattta ctggattgtg gctggtaaag ctattgatta tgaacagatg cttctcctaa
     3001 tgatgaatgt gaagtgggat gtcaaagaga tcatgtcaca gcacaacatc tatgtagatg
     3061 cattgctgaa ggaatttgag cagtttaaca agcggttaaa tgaagtttct aagagagttc
     3121 ggatacctct gcctgtatca aatatacttt gggagcattg tatacgatta gctaacagaa
     3181 ctattgtgga agggtacgcc aatgtcaaga aatgcagtaa tgaggggcgt gccttgatgc
     3241 aattggattt tcaacagttt ttaatgaaac tcgaaaaact aactgacatt agacctattc
     3301 ctgataaaga atttgtggaa acgtatatca aagcctacta cctaactgag aatgacatgg
     3361 agcgttggat caaagagcac agggaatact caacgaagca gttgaccaac ctggtgaatg
     3421 tttgcctagg atctcatatc aataagaaag caagacagaa gctcctagca gctatagatg
     3481 agatagacag gcctaaaaga ccaccggtcg ccaccatggt gagcaagggc gaggagctgt
     3541 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3601 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     3661 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     3721 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3781 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3841 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3901 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3961 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     4021 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     4081 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     4141 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     4201 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat
     4261 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4321 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4381 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4441 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     4501 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4561 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4621 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4681 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4741 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4801 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4861 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4921 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5041 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     5101 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5161 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5221 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5281 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5341 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5401 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     5461 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5521 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5581 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5641 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5701 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5761 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5821 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5881 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5941 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     6001 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6061 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6121 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6181 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6241 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6301 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6361 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6421 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     6481 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6541 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6601 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6661 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     6721 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6781 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6841 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6901 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6961 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7321 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7381 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7441 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7501 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7561 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagt
//
