LOCUS       sequence-373596-        4650 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 a regulatory 
            sequence cloned in the Swa1 restriction site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4650)
  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 4650)
  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
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..41
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        122..140
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     122..139
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    213..337
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(362..367,368..385,386..392)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             446..1105
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(512..531)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1447..1752
                     /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     1631..1650
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1793..1917)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2255..2389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2292..2311)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2346..2365
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2425..2444)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2426..2444)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2531..2548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2702..3290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2782..2801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3461..4252,4253..4321))
                     /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     4084..4103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4322..4426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4494..4512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4550..4572)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
       61 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
      121 gatttaggtg acactataga actcgagcag ctgaagcttg catgcctgca ggtcgactct
      181 agaggatcca tttaaatagg cctttgttta tcacaagttt gtacaaaaaa gctgaacgag
      241 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
      301 cataatactg taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg
      361 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag
      421 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
      481 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
      541 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
      601 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
      661 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
      721 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
      781 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
      841 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
      901 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
      961 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1021 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1081 cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca
     1141 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     1201 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     1261 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     1321 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     1381 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     1441 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     1501 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     1561 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     1621 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     1681 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     1741 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     1801 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     1861 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgat
     1921 aaacttagat atcggaattc cagctgagcg ccggtcgcta ccattaccag ttggtctggt
     1981 gtcaaaaata ataataaccg ggcaggccat gtctgcccgt atttcgcgta aggaaatcca
     2041 ttatgtacta tttaaaaaac acaaactttt ggatgttcgg tttattcttt ttcttttact
     2101 tttttatcat gggagcctac ttcccgtttt tcccgatttg gctacatgac atcaaccata
     2161 tcagcaaaag tgatacgggt attatttttg ccgctatttc tctgttctcg ctattattcc
     2221 aaccgctgtt tggtctgctt tctgacaaac tcggaacttg tttattgcag cttataatgg
     2281 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2341 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg acaaagtcaa
     2401 agcggccatc agatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2461 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2521 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2581 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3121 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4321 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4381 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4441 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4501 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4561 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4621 gtcagggcgc gtcagcgggt gttggcgggt
//
