LOCUS       sequence-348370-        8352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  13Myc tagged VPS51 is expressed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8352)
  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 8352)
  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 VPS51-13Myc
FEATURES             Location/Qualifiers
     source          1..8352
                     /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             3478..3975
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     polyA_signal    4056..4177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4067..4086)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4121..4140
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4363..4818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4450..4469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4660..4681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4935..5292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4935..5126
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4958..4978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5143..5278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5343..6137
                     /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(5397..5416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6007..6026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6201..6249
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6290..6309)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6409..6431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6469..6487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6555..6659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6660..6728,6729..7520)
                     /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(6878..6897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7691..8279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8180..8199
                     /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 gccgcagccg cagcggcagg gcctagtcca
     1141 ggaagcggac caggagactc cccagaagga cccgaagggg aggcaccgga aaggagacgc
     1201 aaggctcacg gtatgctcaa actgtattat gggctgtcag aaggcgaggc tgcagggaga
     1261 ccagctggcc cagaccctct ggacccgacc gatctgaacg gagctcattt tgatcctgaa
     1321 gtttacttgg ataaacttag acgagaatgc cctctcgcac aactgatgga ctcagaaact
     1381 gacatggtgc gacagattag agctctcgac tctgatatgc aaactctcgt ttacgaaaac
     1441 tacaacaagt tcatttctgc aacagacact attaggaaga tgaaaaatga ctttcgcaag
     1501 atggaagatg aaatggatag actggctacg aacatggcgg tgattaccga tttttccgcg
     1561 cgaatatccg cgacactgca agaccgacat gagcgaatta caaaactcgc aggcgtccac
     1621 gccttgctta ggaagctcca gttcctgttt gagctcccaa gtcgcctgac aaaatgtgta
     1681 gagcttggcg cttatggaca agctgttaga taccagggaa gggcacaggc agtactccag
     1741 caataccagc acttgccgtc cttccgcgct atacaagatg attgtcaggt cattactgcc
     1801 cgccttgctc agcagcttag acagaggttc cgggaaggag ggagtggggc accggaacaa
     1861 gcggagtgtg tggagttgtt gttggccctg ggggagcccg ccgaggaact ttgtgaggaa
     1921 tttctggccc acgcgagggg aaggcttgag aaagaattgc gaaacctcga ggctgagttg
     1981 ggaccaagcc ccccagcccc agacgtgctg gaatttactg accacggtgg ctccggattt
     2041 gtgggagggc tttgccaggt cgccgcggcc tatcaagaat tgttcgccgc ccaaggtcca
     2101 gcgggcgccg aaaaacttgc cgcatttgct agacagcttg gatcccgata tttcgcattg
     2161 gtagagagga gactcgcgca agaacaaggt ggtggagaca atagtcttct cgtaagagct
     2221 cttgataggt ttcatcggcg cttgagggca cctggtgccc tgcttgctgc cgccggactt
     2281 gccgacgccg ccactgaaat agtcgagcgg gttgcgcgag agcgcctggg ccatcacctt
     2341 cagggcttga gagcagcgtt cctggggtgt cttacagatg tcaggcaagc actcgccgcc
     2401 ccgagggtcg cgggtaagga aggtcctggt ctcgctgaac tcttggcgaa tgtggcttca
     2461 tcaattcttt cacatattaa ggcaagtctg gctgcggttc acctgtttac ggcaaaggag
     2521 gtaagctttt caaacaagcc ttacttccgg ggtgagttct gcagtcaggg tgtccgtgag
     2581 ggcctcatcg tgggcttcgt ccactctatg tgccagacgg ctcagagctt ctgcgacagc
     2641 cctggggaga aggggggtgc cacaccacct gccctgctcc tgctgctctc ccgcctctgc
     2701 ctggactacg agacggccac catctcctac atcctcactc tcactgatga acagtttctg
     2761 gtgcaggatc agttcccagt gacgcccgtg agcacgctgt gtgcagaggc cagggaaacg
     2821 gcgcggcggc tgctgaccca ctacgtgaag gtgcagggcc tggtcatatc acagatgctg
     2881 cgcaagagcg tggagactcg cgactggctc agcactctgg agccccggaa tgtgcgggcc
     2941 gtcatgaagc gggtggtgga ggataccacc gccatcgacg tgcaggtggg gctcctgtac
     3001 gaagagggtg ttcgcaaggc ccagagcagc gactccagca agaggacttt ctccgtgtac
     3061 agcagctctc ggcagcaggg ccgctacgcc cccagctata cccccagtgc cccgatggac
     3121 accaacctct tgagcaatat ccagaagcta ttctctgaac gtattgatgt gttcagccct
     3181 gtggagttca acaaggtgtc ggtgctgacc ggcatcatca agatcagcct gaagacgctg
     3241 ctggagtgtg tgcggctgcg cacctttggg cgcttcgggc tgcagcaggt gcaagtggac
     3301 tgccactttc tgcagctcta cctgtggcgt tttgtggccg acgaagaact cgtgcacttg
     3361 ctgctggacg aagtggtggc ctctgctgcc ctgcgctgcc cagaccctgt gcccatggag
     3421 cccagtgtgg ttgaggtcat ctgcgagcgc ggcgtcgacc ccgggataat taacggtgaa
     3481 caaaaattaa tctccgagga agacttgaac ggtgaacaaa aattaatctc agaagaagac
     3541 ttgaacggac tcgacggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
     3601 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
     3661 aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag
     3721 ctaatctccg aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac
     3781 ggactcgacg gtgaacaaaa gttgatttct gaagaagatt tgaacggtga acaaaagcta
     3841 atctccgagg aagacttgaa cggtgaacaa aaattaatct cagaagaaga cttgaacgga
     3901 ctcgacggtg aacaaaagtt gatttctgaa gaagatttga acggtgaaca aaagctaatc
     3961 tccgaggaag acttgaacgg tgaacaaaaa ttaatcaatc actagtgatg cggccgcttc
     4021 cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca
     4081 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4141 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4201 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     4261 atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc
     4321 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     4381 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4441 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4501 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4561 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4621 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4681 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4741 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4801 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4861 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     4921 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5041 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5221 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5281 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     5341 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5401 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5461 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5521 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5581 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5641 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5701 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5761 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5821 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5881 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5941 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6001 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6061 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6121 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6181 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6241 ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct
     6301 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6361 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6421 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6481 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6541 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6601 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6661 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6721 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6781 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6841 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6901 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6961 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7021 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7081 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7141 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7201 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7261 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7321 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7381 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7441 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7501 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7561 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7621 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7681 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7741 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7801 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7861 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7921 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7981 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8041 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8101 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8161 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8221 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8281 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     8341 ctcgacagat ct
//
