LOCUS       sequence-373583-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for the expression of a fusion of an N-ter 
            protein of interest with Venus into embryos, by mRNA microinjection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5552)
  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 5552)
  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-RfA-Venus
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     627..646
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        627..645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(834..1547,1548..1550,1551..1553))
                     /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(879..900)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1228..1247
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1487..1508
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    1567..1691
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1732..2037)
                     /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(1834..1853)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2379..3038)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2953..2972
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3092..3098,3099..3116,3117..3122))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3147..3271)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     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
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggtaccg
      661 gccgacttgg ccctgcaggg ggggggnact ttctttttat taggagcaga tacgaatggc
      721 tacattttgg gggacaacat tttgtaaagt gtaagttggt attatgtagc ttagagactc
      781 cattcgggtg ttcttgaggc tggtttagtg gtaaccagat ccgcggccgc gatttacttg
      841 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
      901 tgatcgcgct tctcgttggg gtctttgctc agggcggact ggtagctcag gtagtggttg
      961 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1021 tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc
     1081 tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     1141 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1201 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1261 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1321 tggtcggggt agcgggcgaa gcactgcagg ccgtagccca gggtggtcac gagggtgggc
     1381 cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg
     1441 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     1501 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catatctaag
     1561 tttatcacca ctttgtacaa gaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
     1621 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     1681 cagtcactat ggtcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc
     1741 ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac
     1801 ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct
     1861 ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg
     1921 tgcactggcc agggggatca ccatccgtcg cccgggcgtg tcaataatat cactctgtac
     1981 atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa actgcatttc
     2041 accagcccct gttctcgtca gcaaaagagc cgttcatttc aataaaccgg gcgacctcag
     2101 ccatcccttc ctgattttcc gctttccagc gttcggcacg cagacgacgg gcttcattct
     2161 gcatggttgt gcttaccaga ccggagatat tgacatcata tatgccttga gcaactgata
     2221 gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata gcatacctct ttttgacata
     2281 cttcgggtat acatatcagt atatattctt ataccgcaaa aatcagcgcg caaatacgca
     2341 tactgttatc tggcttttag taagccggat ccacgcgttt acgccccgcc ctgccactca
     2401 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     2461 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     2521 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     2581 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     2641 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     2701 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     2761 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     2821 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     2881 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     2941 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     3001 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     3061 aatctcgacg gatcctaact caaaatccac acattatacg agccggaagc ataaagtgta
     3121 aagcctgggg tgcctaatgc ggccgccata gtgactggat atgttgtgtt ttacagtatt
     3181 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     3241 tttctcgttc agcttttttg tacaaacttg tgataaacaa aggcctgaat 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 gc
//
