LOCUS       sequence-79849-e        7661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Xl-VSP2-GFP fusion in Xenopus oocytes or mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7661)
  AUTHORS   Ratzan WJ, Evsikov AV, Okamura Y, Jaffe LA
  TITLE     Voltage sensitive phosphoinositide phosphatases of Xenopus: their 
            tissue distribution and voltage dependence.
  JOURNAL   J Cell Physiol. 2011 Nov;226(11):2740-6. doi: 10.1002/jcp.22854.
  PUBMED    21618529
REFERENCE   2  (bases 1 to 7661)
  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: "10.1002/jcp"; 
            journalName: "J Cell Physiol"; date: "2011-11"; volume: "226"; 
            issue: "11"; pages: "2740-6"
COMMENT     Sequence Label: pcDNA3-Xl-VSP2 GFP
FEATURES             Location/Qualifiers
     source          1..7661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2452..2461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2458..2460,2461..2463,2464..3177)
                     /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(2458..2460,2461..2463,2464..3174)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2503..2524)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2764..2783)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3111..3132
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3213..3231)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3214..3231)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3251..3268)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3257..3481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3527..3955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3614..3633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3824..3845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3964..3984)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3969..4299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4150..4285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4212..4231
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4366..5160
                     /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(4420..4439)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5030..5049
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5334..5455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5371..5390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5425..5444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5504..5520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5504..5520)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5517..5539)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5528..5544
                     /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(5552..5558,5559..5576,5577..5582))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5597..5618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5735..5752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5906..6494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5986..6005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6665..7456,7457..7525))
                     /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     7288..7307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7526..7630)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 atgtcaagtg taagatacga gcagagggaa gaaccttcca tggttaatgg gtttggcaat
      961 actgaggaga aagtagaaat tgataatggg aagagtgaag tggaggagcc agcaacatgg
     1021 tggaacaaac tgaggaaagt gatttctcca tttgtgatgt cctttggatt cagggttttt
     1081 ggagttgttc tgatcatcgt agactttgtt cttgtaattg tggatctgtc tgtgactgac
     1141 aaaagcagcg gggccaccac ggcaatatca tccatttcat tagccatatc cttcttcttt
     1201 cttattgaca taattctaca tatcttcgtg gaagggttca gtcagtattt cagttccaag
     1261 ctgaacatat ttgatgctgc tattgttatc gtgacgctac tcgtcactct ggtgtacgca
     1321 ttcacagact tcagtggagc tactaatatt ccaagaatgg taaactttct ccgaactttg
     1381 agaattatca tattagtgag aatcctccgc ctggcatctc agaagaggca actcgagaaa
     1441 gtaacaagaa ggatggtctc cgaaaacaaa agacggtatc aaaaagatgg gtttgacctg
     1501 gatctcacac atgttactga tcgtataatt gccatgtcat tcccgtcatc tgggaagcaa
     1561 tcgttttata ggaatcctat acaggaggtt gcaaggtttt tagactccaa gcataaggat
     1621 cactacaaaa tctacaacct gtgcagtgaa aaaggatatg atcctaaata ctttcactat
     1681 cgcgtggagc gagtgtttat agatgatcat aatgtcccag tgttggagga catgctgact
     1741 ttcattgcaa gtgtacgggc gtggatggca gaggatcccc aaaatgttat tgctattcac
     1801 tgtaaaggag gcaaaggacg gacgggcacc atggtttgca cataccttgt tgacagtgac
     1861 cagttcgaga gtgctgtgga aagtttggat tattttggag aaagacggac tgatacaagt
     1921 gtaagcacta agttccaagg tgtagaaaca ccttctcaga gcaggtatgt tgcatactat
     1981 gaagttctga agaacaaatt caaccgccaa cttcctccag agaaacaact caggatcaga
     2041 agtatcagaa tccactccat acaaggtgta ggaagaagca atggcacaga tttaaaggtt
     2101 cagataacaa tgaagaaaga agtaatattc cagtgtatct gtgagacaca ggaaaactgt
     2161 aagctgtttt ttgatgctgg gaacaattcc gtagtcataa gcctggaagg ttgtcctgtt
     2221 atctcgggag acatcaaaat aagatttgag tccagcacaa atcttccaaa aggctacgat
     2281 gactgtgcct tctacttctg gttcaacaca tcctttacag aaaacagcag actctacctt
     2341 tccaaaaatg agctggacaa tccccacaaa gaaaaatctg gaaaaatcta caaagaggat
     2401 tttgctgtgg agttaaactt tgagtctgga cccgggatcc accggccggt cgccaccatg
     2461 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2521 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2581 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2641 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2701 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2761 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2821 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2881 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2941 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3001 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3061 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3121 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     3181 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     3241 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3301 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3361 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3421 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3481 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3541 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3601 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3661 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     3721 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3781 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3841 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     3901 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3961 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4141 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4201 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4261 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4321 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4381 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4441 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4501 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4561 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4621 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4681 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4741 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4801 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4861 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4921 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4981 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5041 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5101 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5161 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5221 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5281 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5461 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5641 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5701 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5761 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6481 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6661 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6721 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6781 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6841 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6901 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6961 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7021 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7081 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7141 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7201 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7261 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7321 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7381 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7441 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7501 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7561 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7621 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
