LOCUS       sequence-44150-e        3756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3756)
  AUTHORS   Najdi R, Proffitt K, Sprowl S, Kaur S, Yu J, Covey TM, Virshup DM, 
            Waterman ML
  TITLE     A uniform human Wnt expression library reveals a shared secretory 
            pathway and unique signaling activities.
  JOURNAL   Differentiation. 2012 Sep;84(2):203-13. doi: 
            10.1016/j.diff.2012.06.004. Epub 2012 Jul 9.
  PUBMED    22784633
REFERENCE   2  (bases 1 to 3756)
  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"; doi: 
            "10.1016/j.diff.2012.06.004"; journalName: "Differentiation"; date: 
            "2012-09"; volume: "84"; issue: "2"; pages: "203-13"
COMMENT     Sequence Label: Wnt10B STOP
FEATURES             Location/Qualifiers
     source          1..3756
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1881..1980)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1997..2016)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1998..2016)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2021..2037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2021..2037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2139..2158)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2150..2959
                     /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(2217..2236)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3106..3694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3595..3614
                     /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 tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat gctggaggag ccccggccgc ggcctccgcc
      721 ctcgggcctc gcgggtctcc tgttcctggc gttgtgcagt cgggctctaa gcaatgagat
      781 tctgggcctg aagttgcctg gcgagccgcc gctgacggcc aacaccgtgt gcttgacgct
      841 gtccggcctg agcaagcggc agctaggcct gtgcctgcgc aaccccgacg tgacggcgtc
      901 cgcgcttcag ggtctgcaca tcgcggtcca cgagtgtcag caccagctgc gcgaccagcg
      961 ctggaactgc tccgcgcttg agggcggcgg ccgcctgccg caccacagcg ccatcctcaa
     1021 gcgcggtttc cgagaaagtg ctttttcctt ctccatgctg gctgctgggg tcatgcacgc
     1081 agtagccacg gcctgcagcc tgggcaagct ggtgagctgt ggctgtggct ggaagggcag
     1141 tggtgagcag gatcggctga gggccaaact gctgcagctg caggcactgt cccgaggcaa
     1201 gagtttcccc cactctctgc ccagccctgg ccctggctca agccccagcc ctggccccca
     1261 ggacacatgg gaatggggtg gctgtaacca tgacatggac tttggagaga agttctctcg
     1321 ggatttcttg gattccaggg aagctccccg ggacatccag gcacgaatgc gaatccacaa
     1381 caacagggtg gggcgccagg tggtaactga aaacctgaag cggaaatgca agtgtcatgg
     1441 cacatcaggc agctgccagt tcaagacatg ctggagggcg gccccagagt tccgggcagt
     1501 gggggcggcg ttgagggagc ggctgggccg ggccatcttc attgataccc acaaccgcaa
     1561 ttctggagcc ttccagcccc gtctgcgtcc ccgtcgcctc tcaggagagc tggtctactt
     1621 tgagaagtct cctgacttct gtgagcgaga ccccactatg ggctccccag ggacaagggg
     1681 ccgggcctgc aacaagacca gccgcctgtt ggatggctgt ggcagcctgt gctgtggccg
     1741 tgggcacaac gtgctccggc agacacgagt tgagcgctgc cattgccgct tccactggtg
     1801 ctgctatgtg ctgtgtgatg agtgcaaggt tacagagtgg gtgaatgtgt gtaagtgaat
     1861 tctgaagggt gggcgcgccg acccagcttt cttgtacaaa gttggcatta taagaaagca
     1921 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1981 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2041 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2101 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2161 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2221 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2281 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2341 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2401 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2461 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2521 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2581 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2641 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2701 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2761 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2821 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2881 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2941 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3001 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3241 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3481 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3721 gttcctggcc ttttgctggc cttttgctca catgtt
//
