LOCUS       sequence-369069-        4680 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 a reg. sequence in the Swa1 restriction site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4680)
  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 4680)
  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::RfA-HA
FEATURES             Location/Qualifiers
     source          1..4680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    79..203
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(228..233,234..251,252..258)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             312..971
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(378..397)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1313..1618
                     /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     1497..1516
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1659..1783)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1797..1823
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2151..2285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2188..2207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2242..2261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2321..2340)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2322..2340)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2427..2444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2598..3186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2678..2697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3357..4148,4149..4217))
                     /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     3980..3999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4218..4322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4390..4408
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4446..4468)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4568..4587
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4668..4680,1..6)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4668..4680,1..5)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tatagaactc gagcagctga agcttgcatg cctgcaggtc gactctagag gatccattta
       61 aataggcctt tgtttatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
      121 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
      181 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
      241 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
      301 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
      361 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
      421 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
      481 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
      541 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
      601 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
      661 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
      721 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
      781 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
      841 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
      901 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
      961 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     1021 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     1081 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     1141 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     1201 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     1261 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     1321 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1381 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     1441 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     1501 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     1561 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     1621 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     1681 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     1741 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtgataaac ttagattacc
     1801 catacgacgt gccagactac gcttgaatcg gaattccagc tgagcgccgg tcgctaccat
     1861 taccagttgg tctggtgtca aaaataataa taaccgggca ggccatgtct gcccgtattt
     1921 cgcgtaagga aatccattat gtactattta aaaaacacaa acttttggat gttcggttta
     1981 ttctttttct tttacttttt tatcatggga gcctacttcc cgtttttccc gatttggcta
     2041 catgacatca accatatcag caaaagtgat acgggtatta tttttgccgc tatttctctg
     2101 ttctcgctat tattccaacc gctgtttggt ctgctttctg acaaactcgg aacttgttta
     2161 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2221 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2281 ggatcgacaa agtcaaagcg gccatcagat ctgccggtct ccctatagtg agtcgtatta
     2341 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2401 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3001 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4321 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     4381 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     4441 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4501 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     4561 aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta
     4621 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
//
