LOCUS       sequence-162294-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ciona VSP (without phosphatase activity) in Xenopus 
            Oocyte.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  AUTHORS   Murata Y, Iwasaki H, Sasaki M, Inaba K, Okamura Y
  TITLE     Phosphoinositide phosphatase activity coupled to an intrinsic 
            voltage sensor.
  JOURNAL   Nature. 2005 Jun 30;435(7046):1239-43. Epub 2005 May 18.
  PUBMED    15902207
REFERENCE   2  (bases 1 to 5701)
  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"; journalName: "Nature."; 
            date: "2005-06-30"; volume: "435(7046)"; pages: "1239-43. Epub 2005 
            May 18"
COMMENT     Sequence Label: pCiVSP(C363S)-SD64TF
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      56..65
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     3'UTR           1827..1952
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     rep_origin      complement(2079..2534)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2216..2237)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2428..2447
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2706..2722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2706..2722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2719..2741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2730..2746
                     /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(2754..2760,2761..2778,2779..2784))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2799..2820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2937..2954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3108..3696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3188..3207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3867..4658,4659..4727))
                     /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     4490..4509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4728..4832)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4900..4918
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4956..4978)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5078..5097
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5311..5330
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5620..5638
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5620..5637
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           5650..5692
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     primer_bind     join(5701,1..16)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg gatccgccac
       61 catggaggga ttcgacggtt cagattttag tcctccagct gatttagttg gcgttgacgg
      121 tgcagtcatg cggaacgtcg ttgacgtcac gataaatggt gacgtcactg ctccgccgaa
      181 agctgcgcca agaaaaagtg aatcggtaaa gaaagttcat tggaatgatg tagaccaagg
      241 accgagtgaa aaaccagaga caagacagga ggaacgaata gatatacccg agatttcagg
      301 tctatggtgg ggcgagaatg aacatggagt ggacgatggg agaatggaga tacctactac
      361 tggtgtaggt cgcgtccagt ttcgtgtccg agcagtgatt gatcatctag ggatgcgagt
      421 ctttggagtc ttcctaattt tcttggacat catcctcatg atcattgatc tcagtcttcc
      481 aggaaaaagt gaatcttcac aatcctttta tgacgggatg gctttggctc tttcttgtta
      541 tttcatgctg gatttaggat taaggatatt tgcctacggg cccaagaatt tcttcaccaa
      601 cccctgggag gttgctgatg gtttgattat cgttgtcaca ttcgtcgtca cgatatttta
      661 cactgtgtta gatgaatacg tgcaagaaac aggagccgat ggtttgggga gattggttgt
      721 gttggcccgt ttgctgcgtg tggttcgatt agcaagaata ttttattccc accaacaaat
      781 gaaggcttca agcagaagaa caatatcaca gaacaagcgt cgatacagga aagacgggtt
      841 tgaccttgac ctcacctacg tcactgacca cgtgattgca atgtcgttcc caagcagtgg
      901 acgacaaagt ttgttccgaa acccgatcgg ggaggtaagc cgtttcttta agacgaagca
      961 cccggacaag tttcgcatct acaacttatg cagcgagagg ggttatgacg aaacaaaatt
     1021 cgacaatcat gtctaccgtg ttatgattga cgaccataat gtaccaacct tagttgatct
     1081 tctaaagttc atagatgacg caaaagtatg gatgacgtca gacccagatc acgtgatagc
     1141 gattcacagt aaaggcggga agggaagaac tggtaccttg gtcagctcgt ggctgcttga
     1201 agatgggaag tttgatacgg caaaggaggc gttagaatac tttggaagca gaagaaccga
     1261 cttcgaagtg ggtgacgttt tccagggagt agagaccgca tcccagatcc gctatgtagg
     1321 ttattttgag aaaatcaaaa agaattacgg aggacaactt cccccgatga agaagttaaa
     1381 ggtaactgga gtcaccatca ctgcgataca aggagtcggt aggggcaatg gttctgattt
     1441 atcaatgcaa atcgtatctg aacggcaaga ggttcttctc tgcaagtttg ctgaaggtta
     1501 taactgtgcg ttacaatacg acgccacgga tgattgtgtt acgtgtgagg tgaaaaactg
     1561 tcctgtgcta gcaggagata ttaaagtcag gttcatgtcg acttcgaaat ctctgccacg
     1621 gggatacgac aactgcccat tttacttttg gttcaacacg tcattggttg aaggagatca
     1681 cgtgacttta aaacgagaag agatcgacaa tccgcataaa aagaaaactt ggaaaatcta
     1741 cagggataat ttcactgtaa aacttacgtt ttcagatgct gaagacattt aggcggccgc
     1801 cgcggtacca gatctggtta ccactaaacc agcctcaaga acacccgaat ggagtctcta
     1861 agctacataa taccaactta cactttacaa aatgttgtcc cccaaaatgt agccattcgt
     1921 atctgctcct aataaaaaga aagtttcttc acgatacata tttgaatgta tttagaaaaa
     1981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgatg cggtgtgaaa
     2041 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     2101 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2161 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2221 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2281 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2341 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2401 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2461 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2521 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2581 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2641 tgggtaacgc casscctgca ggtcgactct agaggatccc cgggcgagct cgaattcgta
     2701 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4861 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4921 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4981 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5041 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     5101 agagtgcacc attcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     5161 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     5221 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     5281 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5341 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctggctagc
     5401 gatgaccctg ctgattggtt cgctgaccat ttccgggtgc gggacggcgt taccagaaac
     5461 tcagaaggtt cgtccaacca aaccgactct gacggcagtt tacgagagag atgatagggt
     5521 ctgcttcagt aagccagatg ctacacaatt aggcttgtac atattgtcgt tagaacgcgg
     5581 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaat
     5641 acaagcttgc ttgttctttt tgcagaagct cagaataaac gctcaacttt ggcagatctc
     5701 t
//
