LOCUS       sequence-373582-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for the expression of a fluorescent fusion of 
            N-ter Venus and a protein of interest into embryos, by mRNA 
            microinjection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  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 5566)
  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-Spe3-Venus-RfA
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..99)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     290..311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     575..597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     589..606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     590..606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gap             674..685
                     /estimated_length=12
     protein_bind    842..966
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1007..1312)
                     /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     complement(1109..1128)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1654..2313)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2228..2247
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2367..2373,2374..2391,2392..2397))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2422..2546)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(2559..3269,3270..3272,3273..3275))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2601..2622)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2950..2969
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3209..3230
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     5'UTR           complement(3300..3342)
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     promoter        complement(3367..3385)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3406..3422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3406..3422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3419..3441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3430..3446
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3454..3460,3461..3478,3479..3484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3499..3520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3637..3654)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3808..4396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3888..3907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4567..5358,5359..5427))
                     /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     5190..5209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5428..5532)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5559..5566,1..448)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
       61 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
      121 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
      181 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
      241 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
      301 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
      361 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
      421 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
      481 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
      541 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
      601 gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccggccg acttggccct
      661 gcaggggggg gggnnnnnnn nnnnnttctt tttattagga gcagatacga atggctacat
      721 tttgggggac aacattttgt aaagtgtaag ttggtattat gtagcttaga gactccattc
      781 gggtgttctt gaggctggtt tagtggtaac cagatccgcg gccgcgatat ctaagtttat
      841 caccactttg tacaagaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
      901 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      961 actatggtcg acctgcagac tggctgtgta taagggagcc tgacatttat attccccaga
     1021 acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt
     1081 ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat
     1141 ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac
     1201 tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca
     1261 caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag
     1321 cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac ctcagccatc
     1381 ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc attctgcatg
     1441 gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac tgatagctgt
     1501 cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg acatacttcg
     1561 ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg
     1621 ttatctggct tttagtaagc cggatccacg cgtttacgcc ccgccctgcc actcatcgca
     1681 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     1741 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     1801 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     1861 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     1921 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     1981 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     2041 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     2101 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     2161 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     2221 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     2281 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     2341 cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc
     2401 tggggtgcct aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2461 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2521 cgttcagctt ttttgtacaa acttgtgata aacaaaggct tgtacagctc gtccatgccg
     2581 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     2641 gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
     2701 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg
     2761 atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata
     2821 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     2881 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     2941 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ttggacgtag
     3001 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
     3061 aagcactgca ggccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg
     3121 ccggtggtgc agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     3181 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     3241 accccggtga acagctcctc gcccttgctc accatttttt ctgcctgaat tcagatctgc
     3301 caaagttgag cgtttattct gagcttctgc aaaaagaaca agcaagctta gctccagctt
     3361 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     3421 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3481 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3541 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3601 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3661 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3721 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3781 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3841 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3901 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3961 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4021 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4081 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4141 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4201 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4261 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4321 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4381 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4441 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4501 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4561 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4621 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4681 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4741 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4801 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4861 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4921 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4981 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5041 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5101 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5161 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5221 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5281 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5341 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5401 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5461 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5521 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccc
//
