LOCUS       sequence-369057-        5253 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 HA tag 
            under control of the pFOG ectodermal regulatory sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5253)
  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 5253)
  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-HA
FEATURES             Location/Qualifiers
     source          1..5253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..110
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        191..209
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     191..208
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    855..979
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1004..1009,1010..1027,1028..1034)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1088..1747
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1154..1173)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2089..2394
                     /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     2273..2292
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2435..2559)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2573..2599
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2927..3061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2964..2983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3018..3037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3097..3116)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3098..3116)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3203..3220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3374..3962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3454..3473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4133..4924,4925..4993))
                     /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     4756..4775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4994..5098)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5166..5184
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5222..5244)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
       61 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      121 accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca
      181 tacacatacg atttaggtga cactatagaa ctcgagcagc tgaagcttgc atgcctgcag
      241 gtcgactcta gaggatccat ttatttctat acatgatggc atcagctagt aagatagaaa
      301 acaaccttgt tattactcgt gctgtaataa tataattcat gaaaaacaaa catggtctta
      361 tcaaatctat tccgaagaca aacatacggc tgatagaatt cagtcagcgc ggtttgggcg
      421 acatcactca gagctctagc tcttatcttg cttatcgagc aagcgataaa acaagcagta
      481 acgagaaaac aagagaaaga aggctagctt tctggagaag accaagataa agtatctcaa
      541 aattcaggaa acggtccaag accgaagctc caaagctctt gtgttcagtt aaactctgat
      601 agtgaataag cttcgtgtat tgtaccggcc cattgtcaat catgcaaact tgatattata
      661 ttgacaagag aagaaggcag tttaaattaa aactctaaag tagagagaca ttaatctcag
      721 ctgacaaggc aggtggtcac agtaagttca tttaaatagt tggccaacaa tagcctttcc
      781 aagaaagtat ttttgttcca ggtctataca aaaataacac acataggatc ccgataaata
      841 ggcctttgtt tatcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
      901 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
      961 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     1021 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
     1081 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1141 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1201 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1261 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     1321 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1381 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1441 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1501 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1561 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1621 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     1681 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1741 ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     1801 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     1861 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     1921 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     1981 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     2041 ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag
     2101 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     2161 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     2221 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     2281 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     2341 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     2401 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     2461 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     2521 attttacgtt tctcgttcag ctttcttgta caaagtggtg ataaacttag attacccata
     2581 cgacgtgcca gactacgctt gaatcggaat tccagctgag cgccggtcgc taccattacc
     2641 agttggtctg gtgtcaaaaa taataataac cgggcaggcc atgtctgccc gtatttcgcg
     2701 taaggaaatc cattatgtac tatttaaaaa acacaaactt ttggatgttc ggtttattct
     2761 ttttctttta cttttttatc atgggagcct acttcccgtt tttcccgatt tggctacatg
     2821 acatcaacca tatcagcaaa agtgatacgg gtattatttt tgccgctatt tctctgttct
     2881 cgctattatt ccaaccgctg tttggtctgc tttctgacaa actcggaact tgtttattgc
     2941 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3001 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     3061 cgacaaagtc aaagcggcca tcagatctgc cggtctccct atagtgagtc gtattaattt
     3121 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3181 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3241 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3301 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3541 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3781 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5101 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5161 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5221 acctctgaca catgcagctc ccggagacgg tca
//
