LOCUS       sequence-373697-        5948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for ascidian transgenesis by electroporation of a
            Gateway-cloned fusion of N-ter Venus and a gene of interest under 
            control of the pFOG ectodermal regulatory sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5948)
  AUTHORS   Roure A, Rothbacher U, Robin F, Kalmar E, Ferone G, Lamy C, Missero 
            C, Mueller F, Lemaire P
  TITLE     A multicassette Gateway vector set for high throughput and 
            comparative analyses in ciona and vertebrate embryos.
  JOURNAL   PLoS One. 2007. 2(9):e916.
  PUBMED    17878951
REFERENCE   2  (bases 1 to 5948)
  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: "PLoS One. 
            2007. 2(9):e916."
COMMENT     Sequence Label: pDEST-pSP172BSSPE-pFOGc::venus-RfA
FEATURES             Location/Qualifiers
     source          1..5948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(646..648,649..651,652..1362)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(691..712)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(952..971)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1299..1320
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1375..1499
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1524..1529,1530..1547,1548..1554)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1608..2267
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1674..1693)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2609..2914
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2793..2812
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2955..3079)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3417..3551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3454..3473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3508..3527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3587..3606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3588..3606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3693..3710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3864..4452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3944..3963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4623..5414,5415..5483))
                     /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     5246..5265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5484..5588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5656..5674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5712..5734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5834..5853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5934..5948,1..4)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5934..5948,1..3)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tagaactcga gcagctgaag cttgcatgcc tgcaggtcga ctctagagga tccatttatt
       61 tctatacatg atggcatcag ctagtaagat agaaaacaac cttgttatta ctcgtgctgt
      121 aataatataa ttcatgaaaa acaaacatgg tcttatcaaa tctattccga agacaaacat
      181 acggctgata gaattcagtc agcgcggttt gggcgacatc actcagagct ctagctctta
      241 tcttgcttat cgagcaagcg ataaaacaag cagtaacgag aaaacaagag aaagaaggct
      301 agctttctgg agaagaccaa gataaagtat ctcaaaattc aggaaacggt ccaagaccga
      361 agctccaaag ctcttgtgtt cagttaaact ctgatagtga ataagcttcg tgtattgtac
      421 cggcccattg tcaatcatgc aaacttgata ttatattgac aagagaagaa ggcagtttaa
      481 attaaaactc taaagtagag agacattaat ctcagctgac aaggcaggtg gtcacagtaa
      541 gttcatttaa atagttggcc aacaatagcc tttccaagaa agtatttttg ttccaggtct
      601 atacaaaaat aacacacata ggatcccgat aaataggcag aaaaaatggt gagcaagggc
      661 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      721 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      781 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      841 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      901 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      961 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1021 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1081 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1141 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1201 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1261 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1321 accgccgccg ggatcactct cggcatggac gagctgtaca agcctttgtt tatcacaagt
     1381 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     1441 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     1501 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     1561 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1621 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1681 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1741 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1801 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1861 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1921 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1981 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     2041 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     2101 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2161 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     2221 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     2281 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     2341 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     2401 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     2461 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     2521 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     2581 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     2641 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     2701 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     2761 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2821 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2881 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2941 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     3001 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     3061 ctttcttgta caaagtggtg ataaacttag atatcggaat tccagctgag cgccggtcgc
     3121 taccattacc agttggtctg gtgtcaaaaa taataataac cgggcaggcc atgtctgccc
     3181 gtatttcgcg taaggaaatc cattatgtac tatttaaaaa acacaaactt ttggatgttc
     3241 ggtttattct ttttctttta cttttttatc atgggagcct acttcccgtt tttcccgatt
     3301 tggctacatg acatcaacca tatcagcaaa agtgatacgg gtattatttt tgccgctatt
     3361 tctctgttct cgctattatt ccaaccgctg tttggtctgc tttctgacaa actcggaact
     3421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3541 atgtctggat cgacaaagtc aaagcggcca tcagatctgc cggtctccct atagtgagtc
     3601 gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga
     3661 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4561 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4741 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5581 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5641 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5701 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5761 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5821 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg
     5881 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg
     5941 tgacacta
//
