LOCUS       sequence-373700-        5905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for ascidian transgenesis by electroporation of a
            Gateway-cloned fusion of a gene of interest with a C-ter ECFP under 
            control of the pFOG ectodermal regulatory sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5905)
  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 5905)
  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::RfA-ECFP
FEATURES             Location/Qualifiers
     source          1..5905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    647..771
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(796..801,802..819,820..826)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             880..1539
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(946..965)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1881..2186
                     /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     2065..2084
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2227..2351)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2367..2369,2370..2882)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(2409..2430)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2670..2689)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3017..3038
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3373..3507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3410..3429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3464..3483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3543..3562)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3544..3562)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3649..3666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3820..4408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3900..3919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4579..5370,5371..5439))
                     /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     5202..5221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5440..5544)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5612..5630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5668..5690)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5790..5809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aactcgagca gctgaagctt gcatgcctgc aggtcgactc tagaggatcc atttatttct
       61 atacatgatg gcatcagcta gtaagataga aaacaacctt gttattactc gtgctgtaat
      121 aatataattc atgaaaaaca aacatggtct tatcaaatct attccgaaga caaacatacg
      181 gctgatagaa ttcagtcagc gcggtttggg cgacatcact cagagctcta gctcttatct
      241 tgcttatcga gcaagcgata aaacaagcag taacgagaaa acaagagaaa gaaggctagc
      301 tttctggaga agaccaagat aaagtatctc aaaattcagg aaacggtcca agaccgaagc
      361 tccaaagctc ttgtgttcag ttaaactctg atagtgaata agcttcgtgt attgtaccgg
      421 cccattgtca atcatgcaaa cttgatatta tattgacaag agaagaaggc agtttaaatt
      481 aaaactctaa agtagagaga cattaatctc agctgacaag gcaggtggtc acagtaagtt
      541 catttaaata gttggccaac aatagccttt ccaagaaagt atttttgttc caggtctata
      601 caaaaataac acacatagga tcccgataaa taggcctttg tttatcacaa gtttgtacaa
      661 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
      721 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
      781 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
      841 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
      901 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
      961 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1021 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1081 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1141 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1201 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1261 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1321 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1381 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1441 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1501 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta
     1561 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     1621 gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga
     1681 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     1741 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     1801 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     1861 cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     1921 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     1981 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     2041 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     2101 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     2161 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     2221 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     2281 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     2341 tacaaagtgg tgataaactt agattggtga gcaagggcga ggagctgttc accggggtgg
     2401 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2461 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2521 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
     2581 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2641 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2701 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2761 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
     2821 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     2881 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2941 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     3001 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccgat cggaattcca
     3061 gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat aataaccggg
     3121 caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt taaaaaacac
     3181 aaacttttgg atgttcggtt tattcttttt cttttacttt tttatcatgg gagcctactt
     3241 cccgtttttc ccgatttggc tacatgacat caaccatatc agcaaaagtg atacgggtat
     3301 tatttttgcc gctatttctc tgttctcgct attattccaa ccgctgtttg gtctgctttc
     3361 tgacaaactc ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3421 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3481 atcaatgtat cttatcatgt ctggatcgac aaagtcaaag cggccatcag atctgccggt
     3541 ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg
     3601 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3661 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3721 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3781 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3841 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3901 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3961 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4021 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4081 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4141 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4201 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4261 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4321 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4381 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4441 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4501 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4561 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4621 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4681 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4741 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4801 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4861 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4921 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4981 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5041 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5101 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5161 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5221 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5281 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5341 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5401 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5461 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5521 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5581 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5641 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5701 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5761 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5821 ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat
     5881 acacatacga tttatgacac tatag
//
