LOCUS       sequence-44137-e        3645 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 3645)
  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 3645)
  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: Wnt3A STOP
FEATURES             Location/Qualifiers
     source          1..3645
                     /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(1770..1869)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1886..1905)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1887..1905)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1910..1926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1910..1926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2028..2047)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2039..2848
                     /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(2106..2125)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2995..3583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3484..3503
                     /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 ggccccactc ggatacttct tactcctctg
      721 cagcctgaag caggctctgg gcagctaccc gatctggtgg tcgctggctg ttgggccaca
      781 gtattcctcc ctgggctcgc agcccatcct gtgtgccagc atcccgggcc tggtccccaa
      841 gcagctccgc ttctgcagga actacgtgga gatcatgccc agcgtggccg agggcatcaa
      901 gattggcatc caggagtgcc agcaccagtt ccgcggccgc cggtggaact gcaccaccgt
      961 ccacgacagc ctggccatct tcgggcccgt gctggacaaa gctaccaggg agtcggcctt
     1021 tgtccacgcc attgcctcag ccggtgtggc ctttgcagtg acacgctcat gtgcagaagg
     1081 cacggccgcc atctgtggct gcagcagccg ccaccagggc tcaccaggca agggctggaa
     1141 gtggggtggc tgtagcgagg acatcgagtt tggtgggatg gtgtctcggg agttcgccga
     1201 cgcccgggag aaccggccag atgcccgctc agccatgaac cgccacaaca acgaggctgg
     1261 gcgccaggcc atcgccagcc acatgcacct caagtgcaag tgccacgggc tgtcgggcag
     1321 ctgcgaggtg aagacatgct ggtggtcgca acccgacttc cgcgccatcg gtgacttcct
     1381 caaggacaag tacgacagcg cctcggagat ggtggtggag aagcaccggg agtcccgcgg
     1441 ctgggtggag accctgcggc cgcgctacac ctacttcaag gtgcccacgg agcgcgacct
     1501 ggtctactac gaggcctcgc ccaacttctg cgagcccaac cctgagacgg gctccttcgg
     1561 cacgcgcgac cgcacctgca acgtcagctc gcacggcatc gacggctgcg acctgctgtg
     1621 ctgcggccgc ggccacaacg cgcgagcgga gcggcgccgg gagaagtgcc gctgcgtgtt
     1681 ccactggtgc tgctacgtca gctgccagga gtgcacgcgc gtctacgacg tgcacacctg
     1741 caagtgaatt ctgaagggtg ggcgcgccga cccagctttc ttgtacaaag ttggcattat
     1801 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1861 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     1921 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     1981 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2041 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2101 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2161 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2221 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2281 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2341 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2401 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2461 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2521 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2581 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2641 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2701 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2761 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2821 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     2881 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     2941 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3001 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3061 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3121 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3361 cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa
     3421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
