LOCUS       sequence-348376-        8000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  V5 tagged VPS52 is expressed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8000)
  AUTHORS   Perez-Victoria FJ, Mardones GA, Bonifacino JS
  TITLE     Requirement of the human GARP complex for mannose 
            6-phosphate-receptor-dependent sorting of cathepsin D to lysosomes.
  JOURNAL   Mol Biol Cell. 2008 Jun;19(6):2350-62. doi: 10.1091/mbc.e07-11-1189.
            Epub 2008 Mar 26.
  PUBMED    18367545
REFERENCE   2  (bases 1 to 8000)
  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: 
            "10.1091/mbc.e07-11-1189"; journalName: "Mol Biol Cell"; date: 
            "2008-06"; volume: "19"; issue: "6"; pages: "2350-62"
COMMENT     Sequence Label: pEF6 VPS52-V5-6His
FEATURES             Location/Qualifiers
     source          1..8000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        212..1390
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          443..1381
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1338..1358
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1407..1426
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1407..1425
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1518..1527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3737..3778
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3788..3805
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3828..3845)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3834..4058
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4104..4532
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4191..4210)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4401..4422
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4541..4561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4546..4875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4726..4861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4788..4807
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4923..4970
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4989..5387
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5545..5666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5582..5601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5636..5655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5715..5731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5715..5731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5728..5750)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5739..5755
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5763..5769,5770..5787,5788..5793))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5808..5829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5946..5963)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6106..6694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6186..6205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6865..7656,7657..7725))
                     /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     7488..7507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7726..7830)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7905..7924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
       61 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctaggctttt
      121 gcaaaaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
      181 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
      241 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      301 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      361 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      421 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      481 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
      541 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
      601 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
      661 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
      721 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
      781 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
      841 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
      901 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
      961 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     1021 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     1081 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     1141 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     1201 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     1261 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     1321 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     1381 ggtgtcgtga ggaattagct tggtactaat acgactcact atagggagac ccaagctggc
     1441 taggtaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat tgcccttggg
     1501 aatggccgcc gctgcgacca tggcggctgc ggcccgggaa ctggtgttgc gggctgggac
     1561 ctcagatatg gaggaggaag agggcccgct ggcgggtggt cctgggctcc aggaaccact
     1621 gcaacttggg gagttggata tcacttctga tgaattcatc ctggatgaag tggatgttca
     1681 cattcaggca aatctggagg atgagttagt aaaggaagct cttaaaacgg gtgtagatct
     1741 ccgtcactat tcaaagcaag ttgagctgga gctacagcag attgaacaga aatccattcg
     1801 ggattatatt caagagagtg agaatatagc atctctacac aaccagatca cagcctgtga
     1861 tgctgtcctg gagcgaatgg agcagatgtt gggagctttt cagagtgacc tcagctccat
     1921 cagctctgag atccggacac tgcaggaaca gtcaggagcc atgaacattc gacttcgaaa
     1981 tcgccaggca gttcggggga aacttgggga gcttgttgat ggtctggtgg tgccttctgc
     2041 tctggtcacg gcaattctgg aggctccagt gacagagccc aggttcttgg agcagctaca
     2101 ggagctggat gccaaggcag ccgcagtcag agagcaggaa gctagaggca cagcagcctg
     2161 cgcagatgtc agaggcgtgc tcgatcggct ccgggtcaag gcagtgacga agatccgaga
     2221 gtttatcctc cagaagattt attccttcag gaaacccatg accaactatc agatccccca
     2281 gacggccctg ctgaagtaca ggttcttcta tcagtttctg ctgggcaatg aacgagcaac
     2341 agcaaaggag atcagggatg aatatgtgga gacgctgagc aagatttact tgtcttacta
     2401 ccgctcttac ctggggcggc tcatgaaggt gcagtatgag gaagtcgctg agaaagatga
     2461 tctaatgggt gtggaagata cagcaaagaa aggattcttc tcaaagccat cgctccgcag
     2521 caggaacacc attttcaccc taggaacccg cggctctgtc atctccccca ctgaacttga
     2581 ggcccccatc ctggtgcctc acacagcgca gcgcggagag cagaggtatc catttgaggc
     2641 cctcttccgc agccagcact acgccctcct agacaattcc tgccgcgaat accttttcat
     2701 ctgtgaattt tttgttgtgt ctggcccagc tgcacacgac ctgttccatg ctgtcatggg
     2761 ccgtacactc agcatgaccc tgaaacacct ggattcttat ctagctgact gctacgatgc
     2821 cattgctgtt tttctctgta tccacattgt tctccggttc cgtaacattg cagcaaagag
     2881 ggatgttcct gccctggaca ggtactggga acaggtgctt gccttgctat ggccacggtt
     2941 tgaactgatc ctggagatga atgttcagag cgtccgaagc actgaccccc agcgcctagg
     3001 ggggttggat actcggcccc actatatcac acgccgctat gcagagttct cctccgctct
     3061 tgtcagtatc aaccagacaa ttcctaatga acggaccatg caattgctgg gacagctgca
     3121 ggtggaggtg gagaattttg tcctccgagt ggcagctgag ttctcctcaa ggaaggagca
     3181 gcttgtgttt ctgatcaaca actatgacat gatgctgggt gtgctgatgg agcgggctgc
     3241 agatgacagc aaagaggttg agagcttcca gcagctgctc aatgctcgga cacaggaatt
     3301 cattgaagag ttgctgtctc ccccttttgg gggtttagtg gcatttgtga aggaggctga
     3361 ggctttgatt gagcgtggac aggctgagcg acttcgaggg gaagaagccc gggtaactca
     3421 gctgatccgt ggctttggta gttcctggaa atcatcagtg gaatctctga gtcaggatgt
     3481 aatgcggagt ttcaccgact tcagaaatgg caccagtatc attcagggag cgctgaccca
     3541 gctgatccag ctctatcatc gcttccaccg ggtgctgtcc cagccgcagc tccgagccct
     3601 ccctgcccgg gctgagctca tcaacattca ccaccttatg gtggagctca agaagcataa
     3661 gcccaacttc aagggcaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     3721 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     3781 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     3841 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3901 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3961 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4021 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4081 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4141 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4201 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4261 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4321 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4381 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4441 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4501 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4681 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4741 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4801 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4861 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     4921 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     4981 actaaaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
     5041 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
     5101 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
     5161 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
     5221 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
     5281 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
     5341 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc
     5401 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5461 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5521 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5581 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5641 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5701 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5761 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5821 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5881 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5941 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6061 tgagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6181 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6301 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6541 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6661 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6721 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6781 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6901 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6961 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7021 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7081 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7141 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7201 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7261 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7321 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7381 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7441 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7501 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7561 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7621 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7681 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7741 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7801 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7861 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     7921 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7981 gcgagcaaaa tttaagctac
//
