LOCUS       sequence-373453-        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for the expression of 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 4876)
  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 4876)
  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
FEATURES             Location/Qualifiers
     source          1..4876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(141..209,210..1001)
                     /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     complement(359..378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1172..1760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1661..1680
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1914..1931
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2048..2069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2084..2089,2090..2107,2108..2114)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2122..2138
                     /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)."
     primer_bind     2127..2149
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2146..2162
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2146..2162
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2183..2201
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           2226..2268
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    2297..2421
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2446..2451,2452..2469,2470..2476)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2530..3189
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2596..2615)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3531..3836
                     /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     3715..3734
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3877..4001)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     3'UTR           4043..4168
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     gap             4171..4202
                     /estimated_length=32
     primer_bind     complement(4246..4265)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4247..4265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4272..4289)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4272..4288)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4281..4303)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(join(4430..4876,1..9))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4567..4588)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4779..4798
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
       61 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      121 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      181 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      241 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      301 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      361 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      421 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      481 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      541 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      601 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      661 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      721 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      781 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      841 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      901 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      961 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1021 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1081 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1141 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1201 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1261 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1321 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1381 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1441 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1501 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1561 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1621 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1681 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1741 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1801 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1861 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1921 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1981 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2041 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2101 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2161 accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggagctaa
     2221 gcttgcttgt tctttttgca gaagctcaga ataaacgctc aactttggca gatctgaatt
     2281 caggcctttg tttatcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
     2341 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     2401 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct
     2461 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
     2521 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     2581 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     2641 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2701 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     2761 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2821 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     2881 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2941 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     3001 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     3061 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     3121 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     3181 ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     3241 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     3301 ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     3361 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     3421 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     3481 gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta
     3541 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     3601 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     3661 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     3721 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     3781 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     3841 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
     3901 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     3961 tcattttacg tttctcgttc agctttcttg tacaaagtgg tgataaactt agatatcgcg
     4021 gccgcggatc tggttaccac taaaccagcc tcaagaacac ccgaatggag tctctaagct
     4081 acataatacc aacttacact ttacaaaatg ttgtccccca aaatgtagcc attcgtatct
     4141 gctcctaata aaaagaaagt ttcttcacat nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     4201 nnymcccccc cccctgcagg gccaagtcgg ccggtaccca attcgcccta tagtgagtcg
     4261 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4321 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4381 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggca aattgtaagc
     4441 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4501 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4561 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4621 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4681 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4741 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4801 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4861 cttaatgcgc cgctac
//
