LOCUS       sequence-373417-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway donor vector with AttP3 AttP4 recombination sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  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 4767)
  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-P4
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(280..307)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(399..485)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     534..556
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     548..565
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     549..565
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    588..819
                     /gene="mutant version of attP"
                     /label=attP3
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1215..1520)
                     /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(1317..1336)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1840..2523)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2438..2457
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2524..2626)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2771..3002)
                     /gene="mutant version of attP"
                     /label=attP4
                     /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 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
       61 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      121 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      181 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      241 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
      301 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
      361 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
      421 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
      481 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc
      541 acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc ctctagaaaa taatgatttt
      601 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat
      661 gccaactttg tataataaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
      721 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      781 actatgaatc aactacttag atggtattag tgacctgtag tcgaccgaca gccttccaaa
      841 tgttcttcgg gtgatgctgc caacttagtc gaccgacagc cttccaaatg ttcttctcaa
      901 acggaatcgt cgtatccagc ctactcgcta ttgtcctcaa tgccgtatta aatcataaaa
      961 agaaataaga aaaagaggtg cgagcctctt ttttgtgtga caaaataaaa acatctacct
     1021 attcatatac gctagtgtca tagtcctgaa aatcatctgc atcaagaaca atttcacaac
     1081 tcttatactt ttctcttaca agtcgttcgg cttcatctgg attttcagcc tctatactta
     1141 ctaaacgtga taaagtttct gtaatttcta ctgtatcgac ctgcagactg gctgtgtata
     1201 agggagcctg acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt
     1261 tcgcggtggc tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata
     1321 tcggtggtca tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg
     1381 gagactttat ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc
     1441 gtgtcaataa tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag
     1501 gtgtaaacct taaactgcat ttcaccagcc cctgttctcg tcagcaaaag agccgttcat
     1561 ttcaataaac cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc
     1621 acgcagacga cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc
     1681 atatatgcct tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc
     1741 atagcatacc tctttttgac atacttcggg tatacatatc agtatatatt cttataccgc
     1801 aaaaatcagc gcgcaaatac gcatactgtt atctggcttt tagtaagccg gatccacgcg
     1861 gcgtttacgc cccccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     1921 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     1981 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     2041 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     2101 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     2161 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     2221 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     2281 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     2341 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     2401 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     2461 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     2521 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     2581 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     2641 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     2701 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     2761 cttagtcgac tacaggtcac taataccatc taagtagttg attcatagtg actggatatg
     2821 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     2881 atttatatca ttttacgttt ctcgttcaac ttttctatac aaagttggca ttataagaaa
     2941 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     3001 ttgccatcca gctgatatcc 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 ggccttt
//
