LOCUS       sequence-79845-e        6913 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression of Xl-VSP1 inactive mutant in Xenopus oocytes or 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6913)
  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 6913)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-VSP1 C301S
FEATURES             Location/Qualifiers
     source          1..6913
                     /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"
     promoter        complement(2465..2483)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2466..2483)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2503..2520)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2509..2733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2779..3207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2866..2885)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3076..3097
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3216..3236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3221..3551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3402..3537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3464..3483
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3618..4412
                     /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(3672..3691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4282..4301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4586..4707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4623..4642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4677..4696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4756..4772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4756..4772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4769..4791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4780..4796
                     /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(4804..4810,4811..4828,4829..4834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4849..4870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4987..5004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5158..5746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5238..5257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5917..6708,6709..6777))
                     /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     6540..6559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6778..6882)
                     /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 taacagacga gcagagggaa gaaccttcca tggttaatgg gttcggcaag
      961 actgaggcga aagtaataat tgataatggg aagaatgaag tggaagagcc agcgacatgg
     1021 tggaacaaac tgaggaaagt aatttctcca tttgtgatgt cctttggatt cagggtcttt
     1081 ggagttgttc tgatcatcgt tgactttgtt ctggtgattg tggatctgtc tgtcattgac
     1141 aaaagcagag aggccaccac tgcaatatca tccatttctt tagccatttc cttcttcttt
     1201 ctcattgatg tgcttctaca tatcttcgta gaagggttcc gtcagtattt cagttccaag
     1261 ctgaacatat ttgatgcggc tattgttatt gtgacactgc ttgtcactct ggtgtatgca
     1321 ttcacagact tcagcggagc tactaatatt ccaagactgg taaactttct tcgaggcttg
     1381 agaattatca tcttggtgag aattctccgc ctggcatctc agaagaggca acttgagaaa
     1441 gtaacaagga ggctggtctc tgaaaacaaa agacgatatc aaaaggatgg gtttgacctg
     1501 gatctcacat atattactga tcgtataatt gccatgtcct ttccatcatc tgggaagcaa
     1561 tcgttttata ggaatccgat tgaggatgta gtaaggtttt tagactccaa gcataaggat
     1621 cactacaaaa tctacaacct gtgcagtgaa aaaggatatg atccaaaata ctttcactac
     1681 cgcgtggagc gagtgtttat agatgatcat aatgtcccag tgctggcgga catgctgact
     1741 ttcactgcga gtgtacgtgc atggatggca gaggatcccc aaaatgttat tgctattcac
     1801 agtaaaggag gcaaaggacg gacggggacc atggtttgca cataccttgt tgacagtgac
     1861 cagtttgaga gtgctgtgga aagtttggat tattttggag aaagacgtac tgataaaagt
     1921 gtaagcacca agttccaggg tgtagaaaca ccttctcaga gcaggtatgt tgcatactat
     1981 gaagttctga agaacaaatt ctgccgccaa cttcctccag agaaacaact caagatcaga
     2041 agtatcaaaa tccactctat acaaggtgta gggagaagca acggcacaga tttaaaagtt
     2101 cagataataa tgaagaaaga agtagtgttc cagtgcgtct gtgagacaca ggaaaactgt
     2161 aagctgtttt ttgatgctgg aaacaactcc atagtcctaa gcctggaagg ttttccagtt
     2221 atctcgggag acattaaaat aagatttgag ttcagcacga atcttccaaa aggctacgat
     2281 gactgtgcca tctacttctg gttcaacaca tgcttcatag aaaacagcag actctacctt
     2341 tccaagaatg agcttgacaa tccccacaaa gaaaaatctg ggaagatcta caaagagaac
     2401 tttgcggtgg agttaaactt tgagtcttga gcggccgctc gagcatgcat ctagagggcc
     2461 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     2521 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2581 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2641 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2701 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2761 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2821 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2881 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     2941 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3001 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3061 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3121 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     3181 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3241 aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3481 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3541 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3601 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3661 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3721 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3781 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3841 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3901 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3961 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4021 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4081 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4141 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4201 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4261 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4321 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4381 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4441 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4501 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4561 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4621 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4681 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4741 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4801 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5341 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5581 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6901 gccacctgac gtc
//
