LOCUS       sequence-373698-        5917 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 Venus under
            control of the pFOG ectodermal regulatory sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5917)
  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 5917)
  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-venus
FEATURES             Location/Qualifiers
     source          1..5917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..28
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     10..27
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    674..798
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(823..828,829..846,847..853)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             907..1566
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(973..992)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1908..2213
                     /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     2092..2111
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2254..2378)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2392..2394,2395..2397,2398..2910)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(2437..2458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2698..2717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3045..3066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3410..3544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3447..3466)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3501..3520
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3580..3599)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3581..3599)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3686..3703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3857..4445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3937..3956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4616..5407,5408..5476))
                     /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     5239..5258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5477..5581)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5649..5667
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5705..5727)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5827..5846
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acacatacga tttaggtgac actatagaac tcgagcagct gaagcttgca tgcctgcagg
       61 tcgactctag aggatccatt tatttctata catgatggca tcagctagta agatagaaaa
      121 caaccttgtt attactcgtg ctgtaataat ataattcatg aaaaacaaac atggtcttat
      181 caaatctatt ccgaagacaa acatacggct gatagaattc agtcagcgcg gtttgggcga
      241 catcactcag agctctagct cttatcttgc ttatcgagca agcgataaaa caagcagtaa
      301 cgagaaaaca agagaaagaa ggctagcttt ctggagaaga ccaagataaa gtatctcaaa
      361 attcaggaaa cggtccaaga ccgaagctcc aaagctcttg tgttcagtta aactctgata
      421 gtgaataagc ttcgtgtatt gtaccggccc attgtcaatc atgcaaactt gatattatat
      481 tgacaagaga agaaggcagt ttaaattaaa actctaaagt agagagacat taatctcagc
      541 tgacaaggca ggtggtcaca gtaagttcat ttaaatagtt ggccaacaat agcctttcca
      601 agaaagtatt tttgttccag gtctatacaa aaataacaca cataggatcc cgataaatag
      661 gcctttgttt atcacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
      721 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
      781 acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc
      841 ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa
      901 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
      961 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1021 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1081 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     1141 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1201 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1261 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1321 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1381 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1441 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1501 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1561 gcgtaaacgc gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
     1621 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
     1681 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     1741 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
     1801 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     1861 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
     1921 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     1981 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     2041 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     2101 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
     2161 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
     2221 gctcccttat acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta
     2281 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     2341 ttttacgttt ctcgttcagc tttcttgtac aaagtggtga taaacttaga tatggtgagc
     2401 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2461 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     2521 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2581 accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     2641 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     2701 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2761 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2821 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     2881 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     2941 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3001 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3061 ttcgtgaccg ccgccgggat cactcatcgg aattccagct gagcgccggt cgctaccatt
     3121 accagttggt ctggtgtcaa aaataataat aaccgggcag gccatgtctg cccgtatttc
     3181 gcgtaaggaa atccattatg tactatttaa aaaacacaaa cttttggatg ttcggtttat
     3241 tctttttctt ttactttttt atcatgggag cctacttccc gtttttcccg atttggctac
     3301 atgacatcaa ccatatcagc aaaagtgata cgggtattat ttttgccgct atttctctgt
     3361 tctcgctatt attccaaccg ctgtttggtc tgctttctga caaactcgga acttgtttat
     3421 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3481 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3541 gatcgacaaa gtcaaagcgg ccatcagatc tgccggtctc cctatagtga gtcgtattaa
     3601 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3661 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4261 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5581 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5641 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5701 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5761 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5821 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     5881 aacgcggcta caattaatac ataaccttat gtatcat
//
