LOCUS       sequence-321952-        7119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human VPS35 cDNA C-terminally tagged with eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  AUTHORS   Halff EF, Szulc BR, Lesept F, Kittler JT
  TITLE     SNX27-Mediated Recycling of Neuroligin-2 Regulates Inhibitory 
            Signaling.
  JOURNAL   Cell Rep. 2019 Nov 26;29(9):2599-2607.e6. doi: 
            10.1016/j.celrep.2019.10.096.
  PUBMED    31775031
REFERENCE   2  (bases 1 to 7119)
  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: 
            "10.1016/j.celrep.2019.10"; journalName: "Cell Rep"; date: 
            "2019-11-26- 26"; volume: "29"; issue: "9"; pages: "2599-2607.e6"
COMMENT     Sequence Label: pDEST-eGFP-C1-VPS35
FEATURES             Location/Qualifiers
     source          1..7119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    626..650
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3039..3063)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3083..3092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3089..3091,3092..3094,3095..3808)
                     /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(3089..3091,3092..3094,3095..3805)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3134..3155)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3395..3414)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3742..3763
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3931..4052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3968..3987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4022..4041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4059..4514)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4196..4217)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4408..4427
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4541..4645
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4647..5004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4647..4838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4670..4690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4855..4990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5039..5833
                     /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             5039..5833
                     /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(5093..5112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5703..5722
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6021..6040)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6065..6112
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6441..7029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6930..6949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      601 gaaccgtcag atccgctagc atcaaacaag tttgtacaaa aaagttggca tgcctacaac
      661 acagcagtcc cctcaggatg agcaggaaaa gctcttggat gaagccatac aggctgtgaa
      721 ggtccagtca ttccaaatga agagatgcct ggacaaaaac aagcttatgg atgctctaaa
      781 acatgcttct aatatgcttg gtgaactccg gacttctatg ttatcaccaa agagttacta
      841 tgaactttat atggccattt ctgatgaact gcactacttg gaggtctacc tgacagatga
      901 gtttgctaaa ggaaggaaag tggcagatct ctacgaactt gtacagtatg ctggaaacat
      961 tatcccaagg ctttaccttt tgatcacagt tggagttgta tatgtcaagt catttcctca
     1021 gtccaggaag gatattttga aagatttggt agaaatgtgc cgtggtgtgc aacatccctt
     1081 gaggggtctg tttcttcgaa attaccttct tcagtgtacc agaaatatct tacctgatga
     1141 aggagagcca acagatgaag aaacaactgg tgacatcagt gattccatgg attttgtact
     1201 gctcaacttt gcagaaatga acaagctctg ggtgcgaatg cagcatcagg gacatagccg
     1261 agatagagaa aaaagagaac gagaaagaca agaactgaga attttagtgg gaacaaattt
     1321 ggtgcgcctc agtcagttgg aaggtgtaaa tgtggaacgt tacaaacaga ttgttttgac
     1381 tggcatattg gagcaagttg taaactgtag ggatgctttg gctcaagaat atctcatgga
     1441 gtgtattatt caggttttcc ctgatgaatt tcacctccag actttgaatc cttttcttcg
     1501 ggcctgtgct gagttacacc agaatgtaaa tgtgaagaac ataatcattg ctttaattga
     1561 tagattagct ttatttgctc accgtgaaga tggacctgga atcccagcgg atattaaact
     1621 ttttgatata ttttcacagc aggtggctac agtgatacag tctagacaag acatgccttc
     1681 agaggatgtt gtatctttac aagtctctct gattaatctt gccatgaaat gttaccctga
     1741 tcgtgtggac tatgttgata aagttctaga aacaacagtg gagatattca ataagctcaa
     1801 ccttgaacat attgctacca gtagtgcagt ttcaaaggaa ctcaccagac ttttgaaaat
     1861 accagttgac acttacaaca atattttaac agtcttgaaa ttaaaacatt ttcacccact
     1921 ctttgagtac tttgactacg agtccagaaa gagcatgagt tgttatgtgc ttagtaatgt
     1981 tctggattat aacacagaaa ttgtctctca agaccaggtg gattccataa tgaatttggt
     2041 atccacgttg attcaagatc agccagatca acctgtagaa gaccctgatc cagaagattt
     2101 tgctgatgag cagagccttg tgggccgctt cattcatctg ctgcgctctg aggaccctga
     2161 ccagcagtac ttgattttga acacagcacg aaaacatttt ggagctggtg gaaatcagcg
     2221 gattcgcttc acactgccac ctttggtatt tgcagcttac cagctggctt ttcgatataa
     2281 agagaattct aaagtggatg acaaatggga aaagaaatgc cagaagattt tttcatttgc
     2341 ccaccagact atcagtgctt tgatcaaagc agagctggca gaattgccct taagactttt
     2401 tcttcaagga gcactagctg ctggggaaat tggttttgaa aatcatgaga cagtcgcata
     2461 tgaattcatg tcccaggcat tttctctgta tgaagatgaa atcagcgatt ccaaagcaca
     2521 gctagctgcc atcaccttga tcattggcac ttttgaaagg atgaagtgct tcagtgaaga
     2581 gaatcatgaa cctctgagga ctcagtgtgc ccttgctgca tccaaacttc taaagaaacc
     2641 tgatcagggc cgagctgtga gcacctgtgc acatctcttc tggtctggca gaaacacgga
     2701 caaaaatggg gaggagcttc acggaggcaa gagggtaatg gagtgcctaa aaaaagctct
     2761 aaaaatagca aatcagtgca tggacccctc tctacaagtg cagcttttta tagaaattct
     2821 gaacagatat atctattttt atgaaaagga aaatgatgcg gtaacaattc aggttttaaa
     2881 ccagcttatc caaaagattc gagaagacct cccgaatctt gaatccagtg aagaaacaga
     2941 gcagattaac aaacattttc ataacacact ggagcatttg cgcttgcggc gggaatcacc
     3001 agaatccgag gggccaattt atgaaggtct catcctttac tcaactttct tgtacaaagt
     3061 ggttcgatgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     3121 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3181 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3241 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     3301 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3361 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3421 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3481 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3541 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     3601 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3661 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     3721 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3781 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     3841 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3901 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3961 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4021 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     4081 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4141 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4201 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4261 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4321 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4381 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4441 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4501 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4621 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4681 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4741 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4801 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4861 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4921 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4981 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5041 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5101 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5161 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5221 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5281 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5341 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5401 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5461 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5521 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5581 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5641 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5701 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5761 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5821 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5881 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5941 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6001 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6061 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6121 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6181 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6241 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6301 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6361 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6421 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6481 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6541 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6601 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6661 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6721 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6781 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6841 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6901 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6961 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7021 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7081 gctcacatgt tctttcctgc gttatcccct gattctgtg
//
