LOCUS       sequence-373415-        4773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway donor vector with AttP3 AttP5 recombination sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4773)
  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 4773)
  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: pDONR-221-P3-P5
FEATURES             Location/Qualifiers
     source          1..4773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(274..301)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(393..479)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     528..550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     542..559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     543..559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    582..813
                     /gene="mutant version of attP"
                     /label=attP3
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1209..1514)
                     /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(1311..1330)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1834..2517)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2432..2451
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2518..2620)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2765..2996)
                     /gene="mutant version of attP"
                     /label=attP5
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3020..3039)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3021..3039)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3044..3060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3044..3060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3162..3181)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3173..3982
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3240..3259)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4129..4717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4618..4637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      241 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      301 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      361 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      421 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      481 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      541 ttgtaaaacg acggccagtc ttaagctcgg gcccctctag aaaataatga ttttattttg
      601 actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac
      661 tttgtataat aaagttgaac gagaaacgta aaatgatata aatatcaata tattaaatta
      721 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg
      781 aatcaactac ttagatggta ttagtgacct gtagtcgacc gacagccttc caaatgttct
      841 tcgggtgatg ctgccaactt agtcgaccga cagccttcca aatgttcttc tcaaacggaa
      901 tcgtcgtatc cagcctactc gctattgtcc tcaatgccgt attaaatcat aaaaagaaat
      961 aagaaaaaga ggtgcgagcc tcttttttgt gtgacaaaat aaaaacatct acctattcat
     1021 atacgctagt gtcatagtcc tgaaaatcat ctgcatcaag aacaatttca caactcttat
     1081 acttttctct tacaagtcgt tcggcttcat ctggattttc agcctctata cttactaaac
     1141 gtgataaagt ttctgtaatt tctactgtat cgacctgcag actggctgtg tataagggag
     1201 cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg
     1261 tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg
     1321 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact
     1381 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca
     1441 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     1501 accttaaact gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat
     1561 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag
     1621 acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga catcatatat
     1681 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca
     1741 tacctctttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat
     1801 cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcca cgcggcgttt
     1861 acgccccccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     1921 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     1981 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2041 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2101 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2161 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2221 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2281 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     2341 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     2401 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     2461 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     2521 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     2581 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     2641 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     2701 atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttagt
     2761 cgactacagg tcactaatac catctaagta gttgattcat agtgactgga tatgttgtgt
     2821 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     2881 atcattttac gtttctcgtt caacttttgt atacaaagtt ggcattataa gaaagcattg
     2941 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3001 tccagctcta gatgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     3061 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3121 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3181 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3241 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3301 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3361 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3421 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3481 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3541 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3601 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3661 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3721 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3781 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3841 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3901 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3961 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     4021 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     4081 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4261 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4741 acggttcctg gccttttgct ggccttttgc tca
//
