LOCUS       sequence-178075-        3678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            WNT1 CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3678)
  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 3678)
  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-WNT1
FEATURES             Location/Qualifiers
     source          1..3678
                     /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(1790..1889)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1919..1938)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1920..1938)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1943..1959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1943..1959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2061..2080)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2072..2881
                     /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(2139..2158)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3028..3616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3517..3536
                     /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 agttgtacct tcaccatggg gctctgggcg ctgttgcctg gctgggtttc tgctacgctg
      721 ctgctggcgc tggccgctct gcccgcagcc ctggctgcca acagcagtgg ccgatggtgg
      781 ggtattgtga acgtagcctc ctccacgaac ctgcttacag actccaagag tctgcaactg
      841 gtactcgagc ccagtctgca gctgttgagc cgcaaacagc ggcgtctgat acgccaaaat
      901 ccggggatcc tgcacagcgt gagtgggggg ctgcagagtg ccgtgcgcga gtgcaagtgg
      961 cagttccgga atcgccgctg gaactgtccc actgctccag ggccccacct cttcggcaag
     1021 atcgtcaacc gaggctgtcg agaaacggcg tttatcttcg ctatcacctc cgccggggtc
     1081 acccattcgg tggcgcgctc ctgctcagaa ggttccatcg aatcctgcac gtgtgactac
     1141 cggcggcgcg gccccggggg ccccgactgg cactgggggg gctgcagcga caacattgac
     1201 ttcggccgcc tcttcggccg ggagttcgtg gactccgggg agaaggggcg ggacctgcgc
     1261 ttcctcatga accttcacaa caacgaggca ggccgtacga ccgtattctc cgagatgcgc
     1321 caggagtgca agtgccacgg gatgtccggc tcatgcacgg tgcgcacgtg ctggatgcgg
     1381 ctgcccacgc tgcgcgccgt gggcgatgtg ctgcgcgacc gcttcgacgg cgcctcgcgc
     1441 gtcctgtacg gcaaccgcgg cagcaaccgc gcttcgcggg cggagctgct gcgcctggag
     1501 ccggaagacc cggcccacaa accgccctcc ccccacgacc tcgtctactt cgagaaatcg
     1561 cccaacttct gcacgtacag cggacgcctg ggcacagcag gcacggcagg gcgcgcctgt
     1621 aacagctcgt cgcccgcgct ggacggctgc gagctgctct gctgcggcag gggccaccgc
     1681 acgcgcacgc agcgcgtcac cgagcgctgc aactgcacct tccactggtg ctgccacgtc
     1741 agctgccgca actgcacgca cacgcgcgta ctgcacgagt gtctgtgaaa cccagctttc
     1801 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     1861 actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc gcgatatccc
     1921 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1981 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2041 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2101 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2161 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2221 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2281 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2341 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2401 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2461 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2521 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2581 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2641 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2701 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2761 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2821 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2881 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2941 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3661 gccttttgct cacatgtt
//
