LOCUS       sequence-44148-e        3659 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 3659)
  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 3659)
  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: Wnt9B STOP
FEATURES             Location/Qualifiers
     source          1..3659
                     /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(1784..1883)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1900..1919)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1901..1919)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1924..1940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1924..1940)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2042..2061)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2053..2862
                     /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(2120..2139)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3009..3597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3498..3517
                     /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 gcgccccccg cccgcgctgg ccctggccgg
      721 gctctgcctg ctggcgctgc ccgccgccgc cgcctcctac ttcggcctga ccgggcggga
      781 agtcctgacg cccttcccag gattgggcac tgcggcagcc ccggcacagg gcggggccca
      841 cctgaagcag tgtgacctgc tgaagctgtc ccggcggcag aagcagctct gccggaggga
      901 gcccggcctg gctgagaccc tgagggatgc tgcgcacctc ggcctgcttg agtgccagtt
      961 tcagttccgg catgagcgct ggaactgtag cctggagggc aggacgggcc tgctcaagag
     1021 aggcttcaaa gagacagctt tcctgtacgc ggtgtcctct gccgccctca cccacaccct
     1081 ggcccgggcc tgcagcgctg ggcgcatgga gcgctgcacc tgtgatgact ctccggggct
     1141 ggagagccgg caggcctggc agtggggcgt gtgcggtgac aacctcaagt acagcaccaa
     1201 gtttctgagc aacttcctgg ggtccaagag aggaaacaag gacctgcggg cacgggcaga
     1261 cgcccacaat acccacgtgg gcatcaaggc tgtgaagagt ggcctcagga ccacgtgtaa
     1321 gtgccatggc gtatcaggct cctgtgccgt gcgcacctgc tggaagcagc tctccccgtt
     1381 ccgtgagacg ggccaggtgc tgaaactgcg ctatgactcg gctgtcaagg tgtccagtgc
     1441 caccaatgag gccttgggcc gcctagagct gtgggcccct gccaggcagg gcagcctcac
     1501 caaaggcctg gccccaaggt ctggggacct ggtgtacatg gaggactcac ccagcttctg
     1561 ccggcccagc aagtactcac ctggcacagc aggtagggtg tgctcccggg aggccagctg
     1621 cagcagcctg tgctgcgggc ggggctatga cacccagagc cgcctggtgg ccttctcctg
     1681 ccactgccag gtgcagtggt gctgctacgt ggagtgccag caatgtgtgc aggaggagct
     1741 tgtgtacacc tgcaagcact gaattctaag ggtgggcgcg ccgacccagc tttcttgtac
     1801 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1861 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1921 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1981 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2041 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2101 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2161 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2221 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2281 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2341 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2401 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2461 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2521 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2581 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2641 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2701 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2761 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2821 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2881 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     2941 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3001 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3061 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3121 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3361 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     3421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
