LOCUS       sequence-348369-        8898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  13Myc tagged VPS50 is expressed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  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 8898)
  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: pCI-neo VPS50-13Myc
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4024..4521
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     polyA_signal    4602..4723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4613..4632)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4667..4686
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4909..5364
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4996..5015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5206..5227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5481..5838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5481..5672
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5504..5524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5689..5824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5889..6683
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5943..5962)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6553..6572
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6747..6795
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6836..6855)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6955..6977
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7015..7033)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7101..7205
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7206..7274,7275..8066)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7424..7443)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8237..8825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8726..8745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt cgccaccatg caaaaaatca aatctctcat gacccgacag
     1141 ggtctgaaaa gccctcaaga aagcctcagt gatcttggtg ccatagagag tctccgggtc
     1201 cctggaaagg aagaattcag ggaacttcga gaacagccaa gtgaccctca agctgaacaa
     1261 gagcttatta atagtattga acaagtatat ttttctgtgg attcatttga tattgttaaa
     1321 tatgagctgg agaagcttcc acctgttctc aatttgcaag aattagaggc gtatagagac
     1381 aaattgaaac aacagcaagc tgcagtatct aaaaaagtgg cagatttaat ccttgaaaaa
     1441 cagcctgctt atgtaaagga acttgaaaga gttacctcat tgcagacagg tcttcaatta
     1501 gctgctgtta tctgtacaaa tgggagaaga cacttgaata ttgcaaagga aggttttact
     1561 caagctagtt taggccttct tgcaaatcaa aggaaacgtc agttgctgat tggacttctg
     1621 aaatctctga gaactataaa aacattgcaa agaacagatg tacggttaag tgaaatgctg
     1681 gaggaggaag attatccagg agctattcag ttgtgccttg aatgtcaaaa agctgccagc
     1741 acttttaaac attacagttg tataagtgaa ctgaattcaa agctgcaaga tactttggaa
     1801 cagattgagg aacagctgga cgtagctctt tccaaaatct gcaagaattt tgacattaac
     1861 cattatacca aggttcaaca agcttatcga cttcttggaa aaacacagac agcaatggat
     1921 caacttcata tgcacttcac ccaagccatt cacaacaccg tgtttcaagt tgttcttggt
     1981 tatgtggaac tatgtgcagg aaacacagac acaaaattcc aaaagctgca atataaggat
     2041 ctctgtacac atgttacacc agacagctat attccatgcc ttgcagacct gtgcaaagca
     2101 ctatgggaag ttatgctcag ctattatagg actatggaat ggcatgaaaa gcatgacaat
     2161 gaggatactg cttcagcttc tgaagggagt aatatgatag gtactgaaga aactaatttt
     2221 gatcgtggct acataaaaaa gaaattagaa catggactta cacgaatatg gcaggatgtt
     2281 cagctaaaag taaaaaccta cttgcttgga actgatttgt ctatattcaa atatgatgat
     2341 ttcatctttg ttttggatat aatcagcagg ttgatgcaag ttggagaaga attttgtggt
     2401 agcaagtctg aagttttaca ggaatctatt agaaaacaaa gtgtcaatta tttcaagaat
     2461 taccatagaa cacggctcga tgaactgaga atgttcttag agaatgagac ttgggaactt
     2521 tgtcctgtta agtcaaattt cagcatcttg caacttcatg aatttaaatt catggaacag
     2581 tctcgctccc catcagtttc acctagtaaa cagccagtct caacttcttc aaaaacagtg
     2641 accttgtttg agcagtactg tagtggtggg aatccatttg aaattcaggc caaccacaaa
     2701 gatgaagaaa cagaagatgt cttagcttct aatgggtatg aatctgatga acaagaaaag
     2761 agtgcctatc aagagtatga cagtgacagt gatgttcctg aggaactcaa acgagactat
     2821 gtggatgagc agacaggaga tggtcctgtg aaaagtgttt ctcgggaaac tctaaaaagc
     2881 aggaagaaat cagattacag tctaaataaa gtgaatgcac ctatcttaac aaatacaaca
     2941 ttgaacgtca taagacttgt tggaaaatat atgcagatga tgaacattct taagccaatt
     3001 gcctttgatg ttattcattt catgtctcaa ctatttgatt attacttgta tgcaatatat
     3061 accttttttg gtcggaatga ttcattggaa tcaactggac tcggccttag tagtagtaga
     3121 ctaagaacaa ctctaaacag aatacaagaa agccttattg atctagaagt ttcagctgat
     3181 cctactgcca cactcacagc agcagaagaa agaaaggaga aggtgccaag tccacacctc
     3241 agtcacctag tggttttgac atctggggat acgctgtatg ggttggcaga aagagtggta
     3301 gccacggaat ccttggtatt cttggctgaa cagtttgagt tccttcagcc acatctggat
     3361 gctgtgatgc ctgcagtcaa aaagcccttt cttcagcagt tctattctca gacagtctca
     3421 accgccagtg aactacggaa accaatttac tggattgtag ctggtaaagc ccttgattat
     3481 gaacagatgc tgcttctcat ggctaatgtg aaatgggatg taaaagaaat tatgtcacag
     3541 cacaacatat atgtagatgc actattaaag gaatttgagc agtttaacag gaggctaaat
     3601 gaagtttcta agagagttcg catacccttg cctgtgtcta atatactttg ggaacattgt
     3661 atacgattgg ctaatcgaac tattgtagaa ggatatgcca atgtcaagaa atgcagtaat
     3721 gagggtcgtg ccctgatgca attggatttt caacagtttt taatgaaact tgaaaaacta
     3781 acagatatta gacccattcc tgataaagaa tttgtagaaa cttatattaa agcttattac
     3841 ctaactgaga atgacatgga acggtggatc aaagagcaca gggaatattc aacgaagcag
     3901 ctgaccaatc tggtgaatgt ttgcctggga tcccatatca ataagaaagc aagacaaaaa
     3961 cttctagcag ctatagatga tatagacaga cctaaaagag tcgaccccgg gataattaac
     4021 ggtgaacaaa aattaatctc cgaggaagac ttgaacggtg aacaaaaatt aatctcagaa
     4081 gaagacttga acggactcga cggtgaacaa aagttgattt ctgaagaaga tttgaacggt
     4141 gaacaaaagc taatctccga ggaagacttg aacggtgaac aaaaattaat ctcagaagaa
     4201 gacttgaacg gactcgacgg tgaacaaaag ttgatttctg aagaagattt gaacggtgaa
     4261 caaaagctaa tctccgagga agacttgaac ggtgaacaaa aattaatctc agaagaagac
     4321 ttgaacggac tcgacggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
     4381 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
     4441 aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag
     4501 ctaatctccg aggaagactt gaacggtgaa caaaaattaa tcaatcacta gtgatgcggc
     4561 cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca ttgatgagtt
     4621 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4681 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4741 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     4801 ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag cgaagaggcc
     4861 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     4921 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4981 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5041 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5101 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5161 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5221 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5281 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5341 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     5401 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     5461 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5521 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5581 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5641 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5701 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5761 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5821 ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta
     5881 gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5941 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6001 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6061 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6121 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6181 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6241 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6301 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6361 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6421 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6481 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6541 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     6601 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6661 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     6721 gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     6781 ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa
     6841 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6901 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6961 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     7021 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     7081 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7141 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7201 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7261 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7321 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7381 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7441 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7501 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7561 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7621 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7681 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7741 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7801 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7861 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7921 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7981 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8041 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8101 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8161 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8221 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8281 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8341 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8401 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8461 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8521 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8581 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8641 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8701 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8761 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8821 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8881 acatggctcg acagatct
//
