LOCUS       sequence-178175-        3616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            WNT3A CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3616)
  AUTHORS   Jackel C, Nogueira MS, Ehni N, Kraus C, Ranke J, Dohmann M, Noessner
            E, Nelson PJ
  TITLE     A vector platform for the rapid and efficient engineering of stable 
            complex transgenes.
  JOURNAL   Sci Rep. 2016 Oct 3;6:34365. doi: 10.1038/srep34365.
  PUBMED    27694838
REFERENCE   2  (bases 1 to 3616)
  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: "Sci Rep."; 
            date: "2016-10-3"; volume: "6:34365. doi"; pages: " 
            10.1038/srep34365"
COMMENT     Sequence Label: pENTR221-WNT3A
FEATURES             Location/Qualifiers
     source          1..3616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1728..1827)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1857..1876)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1858..1876)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1881..1897)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1881..1897)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1999..2018)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2010..2819
                     /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(2077..2096)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2966..3554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3455..3474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttgtaatg gccccactcg gatacttctt actcctctgc agcctgaagc aggctctggg
      721 cagctacccg atctggtggt cgctggctgt tgggccacag tattcctccc tgggctcgca
      781 gcccatcctg tgtgccagca tcccgggcct ggtccccaag cagctccgct tctgcaggaa
      841 ctacgtggag atcatgccca gcgtggccga gggcatcaag attggcatcc aggagtgcca
      901 gcaccagttc cgcggccgcc ggtggaactg caccaccgtc cacgacagcc tggccatctt
      961 cgggcccgtg ctggacaaag ctaccaggga gtcggccttt gtccacgcca ttgcctcagc
     1021 cggtgtggcc tttgcagtga cacgctcatg tgcagaaggc acggccgcca tctgtggctg
     1081 cagcagccgc caccagggct caccaggcaa gggctggaag tggggtggct gtagcgagga
     1141 catcgagttt ggtgggatgg tgtctcggga gttcgccgac gcccgggaga accggccaga
     1201 tgcccgctca gccatgaacc gccacaacaa cgaggctggg cgccaggcca tcgccagcca
     1261 catgcacctc aagtgcaagt gccacgggct gtcgggcagc tgcgaggtga agacatgctg
     1321 gtggtcgcaa cccgacttcc gcgccatcgg tgacttcctc aaggacaagt acgacagcgc
     1381 ctcggagatg gtggtggaga agcaccggga gtcccgcggc tgggtggaga ccctgcggcc
     1441 gcgctacacc tacttcaagg tgcccacgga gcgcgacctg gtctactacg aggcctcgcc
     1501 caacttctgc gagcccaacc ctgagacggg ctccttcggc acgcgcgacc gcacctgcaa
     1561 cgtcagctcg cacggcatcg acggctgcga cctgctgtgc tgcggccgcg gccacaacgc
     1621 gcgagcggag cggcgccggg agaagtgccg ctgcgtgttc cactggtgct gctacgtcag
     1681 ctgccaggag tgcacgcgcg tctacgacgt gcacacctgc aagtagaacc cagctttctt
     1741 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1801 tatcagtcaa aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct
     1861 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1921 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1981 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     2041 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     2101 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     2161 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2221 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2281 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     2341 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2401 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2461 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2521 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2581 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2641 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2701 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2761 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2821 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2881 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2941 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3001 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3061 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3421 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3481 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3541 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3601 cttttgctca catgtt
//
