LOCUS       sequence-162392-        5037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ciona VSP in Xenopus Oocyte.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5037)
  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 5037)
  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-SD64TF
FEATURES             Location/Qualifiers
     source          1..5037
                     /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"
     primer_bind     1964..1983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2042..2058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2042..2058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2055..2077)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2066..2082
                     /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(2090..2096,2097..2114,2115..2120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2135..2156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2273..2290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2444..3032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2524..2543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3203..3994,3995..4063))
                     /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     3826..3845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4064..4168)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4236..4254
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4292..4314)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4414..4433
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4647..4666
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4956..4974
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4956..4973
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           4986..5028
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     primer_bind     join(5037,1..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 gattcactgt 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 aagtytsttc acgcattcta gttgtggttt gtccaaactc
     1981 atcaatgtay cytrymggtc gactctagag gatccccggg cgagctcgaa ttcgtaatca
     2041 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2881 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4141 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4201 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4261 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4321 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4381 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4441 tgcaccattc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     4501 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     4561 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     4621 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     4681 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg gctagcgatg
     4741 accctgctga ttggttcgct gaccatttcc gggtgcggga cggcgttacc agaaactcag
     4801 aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga tagggtctgc
     4861 ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga acgcggctac
     4921 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaatacaa
     4981 gcttgcttgt tctttttgca gaagctcaga ataaacgctc aactttggca gatctct
//
