LOCUS       sequence-373599-        5223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for ascidian transgenesis by electroporation of a
            Gateway-cloned 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 5223)
  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 5223)
  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
FEATURES             Location/Qualifiers
     source          1..5223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    655..779
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(804..809,810..827,828..834)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             888..1547
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(954..973)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1889..2194
                     /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     2073..2092
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2235..2359)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2697..2831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2734..2753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2788..2807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2867..2886)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2868..2886)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2973..2990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3144..3732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3224..3243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3903..4694,4695..4763))
                     /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     4526..4545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4764..4868)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4936..4954
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4992..5014)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5114..5133
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5214..5223,1..9)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5214..5223,1..8)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cactatagaa ctcgagcagc tgaagcttgc atgcctgcag gtcgactcta gaggatccat
       61 ttatttctat acatgatggc atcagctagt aagatagaaa acaaccttgt tattactcgt
      121 gctgtaataa tataattcat gaaaaacaaa catggtctta tcaaatctat tccgaagaca
      181 aacatacggc tgatagaatt cagtcagcgc ggtttgggcg acatcactca gagctctagc
      241 tcttatcttg cttatcgagc aagcgataaa acaagcagta acgagaaaac aagagaaaga
      301 aggctagctt tctggagaag accaagataa agtatctcaa aattcaggaa acggtccaag
      361 accgaagctc caaagctctt gtgttcagtt aaactctgat agtgaataag cttcgtgtat
      421 tgtaccggcc cattgtcaat catgcaaact tgatattata ttgacaagag aagaaggcag
      481 tttaaattaa aactctaaag tagagagaca ttaatctcag ctgacaaggc aggtggtcac
      541 agtaagttca tttaaatagt tggccaacaa tagcctttcc aagaaagtat ttttgttcca
      601 ggtctataca aaaataacac acataggatc ccgataaata ggcctttgtt tatcacaagt
      661 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
      721 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
      781 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
      841 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
      901 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
      961 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1021 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1081 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1141 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1201 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1261 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1321 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1381 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1441 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1501 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     1561 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     1621 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     1681 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     1741 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     1801 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     1861 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     1921 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     1981 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     2041 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2101 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2161 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2221 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     2281 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     2341 ctttcttgta caaagtggtg ataaacttag atatcggaat tccagctgag cgccggtcgc
     2401 taccattacc agttggtctg gtgtcaaaaa taataataac cgggcaggcc atgtctgccc
     2461 gtatttcgcg taaggaaatc cattatgtac tatttaaaaa acacaaactt ttggatgttc
     2521 ggtttattct ttttctttta cttttttatc atgggagcct acttcccgtt tttcccgatt
     2581 tggctacatg acatcaacca tatcagcaaa agtgatacgg gtattatttt tgccgctatt
     2641 tctctgttct cgctattatt ccaaccgctg tttggtctgc tttctgacaa actcggaact
     2701 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2761 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2821 atgtctggat cgacaaagtc aaagcggcca tcagatctgc cggtctccct atagtgagtc
     2881 gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga
     2941 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3001 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3061 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3121 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3181 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3241 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3301 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3361 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3421 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3481 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3541 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3601 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3661 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3721 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3781 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3841 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3901 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3961 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4021 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4081 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4141 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4201 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4261 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4321 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4381 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4441 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4501 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4561 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4621 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4681 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4741 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4801 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4861 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4921 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     4981 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5041 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5101 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg
     5161 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg
     5221 tga
//
