LOCUS       sequence-373600-        5375 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 a reg. sequence in the Swa1 restriction site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5375)
  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 5375)
  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-Swa1::venus-RfA
FEATURES             Location/Qualifiers
     source          1..5375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(63..65,66..68,69..779)
                     /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(108..129)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(369..388)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     716..737
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    792..916
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(941..946,947..964,965..971)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1025..1684
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1091..1110)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2026..2331
                     /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     2210..2229
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2372..2496)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2834..2968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2871..2890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2925..2944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3004..3023)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3005..3023)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3110..3127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3281..3869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3361..3380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4040..4831,4832..4900))
                     /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     4663..4682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4901..5005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5073..5091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5129..5151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5251..5270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5351..5369
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5351..5368
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gcagctgaag cttgcatgcc tgcaggtcga ctctagagga tccatttaaa taggcagaaa
       61 aaatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      121 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      181 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
      241 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
      301 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      361 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      421 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      481 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
      541 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
      601 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      661 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
      721 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc
      781 ctttgtttat cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata
      841 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
      901 atatccagtc actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg
      961 ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc
     1021 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1081 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     1141 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1201 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     1261 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1321 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1381 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1441 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1501 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     1561 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     1621 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     1681 gtaaacgcgt ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga
     1741 tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat
     1801 gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca
     1861 tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc
     1921 tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta
     1981 ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt
     2041 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
     2101 gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
     2161 gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
     2221 accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac
     2281 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     2341 tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca
     2401 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
     2461 ttacgtttct cgttcagctt tcttgtacaa agtggtgata aacttagata tcggaattcc
     2521 agctgagcgc cggtcgctac cattaccagt tggtctggtg tcaaaaataa taataaccgg
     2581 gcaggccatg tctgcccgta tttcgcgtaa ggaaatccat tatgtactat ttaaaaaaca
     2641 caaacttttg gatgttcggt ttattctttt tcttttactt ttttatcatg ggagcctact
     2701 tcccgttttt cccgatttgg ctacatgaca tcaaccatat cagcaaaagt gatacgggta
     2761 ttatttttgc cgctatttct ctgttctcgc tattattcca accgctgttt ggtctgcttt
     2821 ctgacaaact cggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2881 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2941 catcaatgta tcttatcatg tctggatcga caaagtcaaa gcggccatca gatctgccgg
     3001 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
     3061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5101 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5161 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5221 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5281 accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca
     5341 tacacatacg atttaggtga cactatagaa ctcga
//
