LOCUS       sequence-348375-        9057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  13Myc tagged VPS54 is expressed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9057)
  AUTHORS   Gershlick DC, Ishida M, Jones JR, Bellomo A, Bonifacino JS, Everman 
            DB
  TITLE     A neurodevelopmental disorder caused by mutations in the VPS51 
            subunit of the GARP and EARP complexes.
  JOURNAL   Hum Mol Genet. 2019 May 1;28(9):1548-1560. doi: 10.1093/hmg/ddy423.
  PUBMED    30624672
REFERENCE   2  (bases 1 to 9057)
  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"; journalName: 
            "Hum Mol Genet"; date: "2019-05-1- 1"; volume: "28"; issue: "9"; 
            pages: "1548-1560"
COMMENT     Sequence Label: pCI-neo VPS54-13Myc
FEATURES             Location/Qualifiers
     source          1..9057
                     /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"
     regulatory      1102..1111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4183..4680
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     polyA_signal    4761..4882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4772..4791)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4826..4845
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5068..5523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5155..5174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5365..5386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5640..5997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5640..5831
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5663..5683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5848..5983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6048..6842
                     /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(6102..6121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6712..6731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6906..6954
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6995..7014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7114..7136
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7174..7192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7260..7364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7365..7433,7434..8225)
                     /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(7583..7602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8396..8984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8885..8904
                     /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 gcttcaagcc acagttcttc accagtgcct
     1141 caaggaagca gcagtgatgt tttctttaaa atagaggtag atccgtcaaa acacattcga
     1201 cctgtgccat cactgccaga tgtgtgtccc aaggaaccca caggtgattc acatagttta
     1261 tatgttgccc catctctagt tacagatcaa catagatgga ctgtatatca ttccaaagta
     1321 aatctcccag cagcattaaa cgatcctaga ttagcaaaaa gagaatctga cttcttcaca
     1381 aaaacatggg gattggactt tgtggacact gaagtcatac cttcattcta cctcccacag
     1441 atcagcaagg aacattttac agtatatcaa caggaaatct ctcagagaga gaagattcat
     1501 gagagatgca agaatatttg tcctcctaaa gataccttcg aaaggactct tttacatact
     1561 catgataaat ccaggacaga tctggagcaa gtacctaaga tttttatgaa accagatttt
     1621 gccttggatg attccttaac ttttaattca gttttaccat ggtctcattt taatactgct
     1681 ggtggaaaag gaaatcgtga tgcagcttcc tcaaagttgc ttcaagaaaa gctgagccat
     1741 tatctggata ttgtggaagt aaacattgct caccagatct ctctacgttc agaagcattt
     1801 tttcatgcaa tgacctctca acacgagttg caggactacc tcaggaaaac ttcccaggct
     1861 gtaaaaatgc ttcgagataa aattgcacag attgataaag taatgtgtga aggatcactc
     1921 cacattttaa gactggcact taccagaaat aattgtgtta aagtatacaa taagctgaag
     1981 ttaatggcca ctgtacacca gactcagcct acagtacagg tgttattatc tacttctgaa
     2041 tttgttggag cattggactt aatagcaaca acacaagagg ttctacagca ggaacttcag
     2101 ggcattcaca gtttccggca tttgggatca cagctttgtg aattagaaaa actgatagat
     2161 aaaatgatga ttgcagaatt ttctacttat tctcacagtg acttaaatag accactggaa
     2221 gatgactgtc aagttttaga agaggaaaga ctaatatctc ttgtatttgg acttttaaaa
     2281 caaagaaagc ttaatttttt agaaatctat ggtgaaaaaa tggttattac agcaaagaat
     2341 atcattaaac agtgtgtgat taataaagtt tcacaaacag aagaaataga cacagatgtt
     2401 gttgtgaagc ttgcagatca gatgagaatg ttgaattttc cccagtggtt tgatctgctc
     2461 aaggatattt tctctaagtt tacaattttc ctacagagag tgaaggcaac attaaatatc
     2521 attcacagtg ttgttctctc agttcttgac aaaaaccaaa ggactagaga attggaagag
     2581 atttcacaac agaagaatgc tgcaaaagat aattcactgg acacagaggt ggcttattta
     2641 atccatgaag gcatgtttat aagtgatgca ttcggtgagg gtgagctaac acctatagca
     2701 gttgacacta cctctcaaag aaatgcatct ccaaatagtg agccctgcag cagtgattct
     2761 gtatccgagc cagaatgtac tactgattct tcatccagca aagagcacac atcatcatct
     2821 gctattccag gaggtgtgga tattatggtc agtgaagata tgaaattaac tgactcagag
     2881 ctaggaaagc tggcaaataa tatccaggaa ttattatata gtgcctcaga tatatgccat
     2941 gatcgagctg tcaaatttct catgtcaaga gcaaaggatg gttttcttga gaagctaaat
     3001 tccatggaat tcataacact ttctagatta atggaaacat tcattttaga caccgaacag
     3061 atctgtggaa gaaaaagcac gtcattactt ggagcacttc agagccaagc tattaagttt
     3121 gtaaataggt ttcatgaaga gagaaaaacc aagctcagcc tcctcttaga caatgagcgc
     3181 tggaagcaag cagatgttcc tgcagaattt caggatcttg ttgattctct gtcagatggg
     3241 aagattgctt tacctgaaaa aaaatcagga gccacagaag aaaggaaacc agctgaagtt
     3301 cttattgtcg agggacaaca gtatgcagtt gttggaaccg tattgctgtt aataagaatt
     3361 atccttgaat attgccagtg tgtggataac atcccatctg ttactactga catgcttact
     3421 cgtctgtcag atttattgaa gtacttcaat tcaagaagtt gccagttagt tcttggagct
     3481 ggtgcactgc aagttgttgg actaaaaacg ataactacaa aaaatttggc tctttcttca
     3541 cgatgtttgc agttaattgt gcactacatt cctgtgatcc gggctcattt tgaagctcga
     3601 ctaccaccta agcaatatag catgcttagg cattttgatc atatcactaa ggactaccat
     3661 gatcacatag ctgaaatatc agctaagctt gtagcgataa tggatagctt atttgacaag
     3721 ctgttatcta agtatgaagt gaaggctcct gttccttctg cctgtttcag gaatatttgt
     3781 aagcaaatga caaaaatgca cgaagctata tttgatctcc ttccagaaga acaaacacag
     3841 atgttatttt taagaattaa tgcaagttat aaactccact tgaaaaagca gttatctcac
     3901 ttaaatgtga taaatgatgg aggacctcaa aatggacagg gtctcatggt cacacaggct
     3961 ggagggcagt ggtgtgatca tggctcactg cagcctcaac ctccgtggct caagcaatcc
     4021 tcccactgca gactccccaa taactgcgac tacaggttgg tcacagcaga tgtagctttt
     4081 tacactggaa atcttcaagc cttaaaaggc cttaaagatt tggacctaaa tatggccgaa
     4141 atttgggagc agaagagggt cgaccccggg ataattaacg gtgaacaaaa attaatctcc
     4201 gaggaagact tgaacggtga acaaaaatta atctcagaag aagacttgaa cggactcgac
     4261 ggtgaacaaa agttgatttc tgaagaagat ttgaacggtg aacaaaagct aatctccgag
     4321 gaagacttga acggtgaaca aaaattaatc tcagaagaag acttgaacgg actcgacggt
     4381 gaacaaaagt tgatttctga agaagatttg aacggtgaac aaaagctaat ctccgaggaa
     4441 gacttgaacg gtgaacaaaa attaatctca gaagaagact tgaacggact cgacggtgaa
     4501 caaaagttga tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac
     4561 ttgaacggtg aacaaaaatt aatctcagaa gaagacttga acggactcga cggtgaacaa
     4621 aagttgattt ctgaagaaga tttgaacggt gaacaaaagc taatctccga ggaagacttg
     4681 aacggtgaac aaaaattaat caatcactag tgatgcggcc gcttcccttt agtgagggtt
     4741 aatgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4801 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4861 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4921 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg
     4981 ataaggatcg atccgggctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5041 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
     5101 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5161 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5221 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5281 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5341 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5401 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5461 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5521 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg
     5581 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc
     5641 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5821 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5881 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5941 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6001 cttgattctt ctgacacaac agtctcgaac ttaaggctag agccaccatg attgaacaag
     6061 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     6121 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     6181 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     6241 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6301 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6361 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6421 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6481 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6541 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     6601 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6661 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6721 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6781 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6841 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg
     6901 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     6961 tagcgataag gatccgcgta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7021 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7081 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7141 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     7201 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     7261 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7321 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     7381 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7441 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7501 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7561 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7621 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7681 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7741 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7801 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7861 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7921 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7981 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8041 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     8101 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8161 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8221 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8281 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     8341 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8401 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8461 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8521 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8581 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8641 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8701 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8761 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8821 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8881 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8941 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9001 cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga cagatct
//
