LOCUS       sequence-178073-        3625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            WNT3 CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3625)
  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 3625)
  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-WNT3
FEATURES             Location/Qualifiers
     source          1..3625
                     /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(1737..1836)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1866..1885)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1867..1885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1890..1906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1890..1906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2008..2027)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2019..2828
                     /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(2086..2105)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2975..3563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3464..3483
                     /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 gagccccacc tgctcgggct gctcctcggc ctcctgctcg gtggcaccag
      721 ggtcctcgct ggctacccaa tttggtggtc cctggccctg ggccagcagt acacatctct
      781 gggctcacag cccctgctct gcggctccat cccaggcctg gtccccaagc aactgcgctt
      841 ctgccgcaat tacatcgaga tcatgcccag cgtggccgag ggcgtgaagc tgggcatcca
      901 ggagtgccag caccagttcc ggggccgccg ctggaactgc accaccatag atgacagcct
      961 ggccatcttt gggcccgtcc tcgacaaagc cacccgcgag tcggccttcg ttcacgccat
     1021 cgcctcggcc ggcgtggcct tcgccgtcac ccgctcctgc gccgagggca cctccaccat
     1081 ttgcggctgt gactcgcatc ataaggggcc gcctggcgaa ggctggaagt ggggcggctg
     1141 cagcgaggac gctgacttcg gcgtgttagt gtccagggag ttcgcggatg cgcgcgagaa
     1201 caggccggac gcgcgctcgg ccatgaacaa gcacaacaac gaggcgggcc gcacgactat
     1261 cctggaccac atgcacctca aatgcaagtg ccacgggctg tcgggcagct gtgaggtgaa
     1321 gacctgctgg tgggcgcagc ctgacttccg tgccatcggt gacttcctca aggacaagta
     1381 tgacagcgcc tcggagatgg tagtagagaa gcaccgtgag tcccgaggct gggtggagac
     1441 cctccgggcc aagtactcgc tcttcaagcc acccacggag agggacctgg tctactacga
     1501 gaactccccc aacttttgtg agcccaaccc agagacgggt tcctttggca caagggaccg
     1561 gacttgcaat gtcacctccc acggcatcga tggctgcgat ctgctctgct gtggccgggg
     1621 ccacaacacg aggacggaga agcggaagga aaaatgccac tgcatcttcc actggtgctg
     1681 ctacgtcagc tgccaggagt gtattcgcat ctacgacgtg cacacctgca agtagaaccc
     1741 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1801 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ggcggccgcg
     1861 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1921 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1981 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     2041 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     2101 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     2161 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     2221 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2281 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     2341 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     2401 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     2461 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     2521 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     2581 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2641 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2701 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2761 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2821 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     2881 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3121 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3601 tttgctggcc ttttgctcac atgtt
//
