LOCUS       sequence-44147-e        3684 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 3684)
  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 3684)
  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: Wnt9A STOP
FEATURES             Location/Qualifiers
     source          1..3684
                     /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(1809..1908)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1925..1944)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1926..1944)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1949..1965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1949..1965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2067..2086)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2078..2887
                     /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(2145..2164)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3034..3622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3523..3542
                     /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 gctggatggg tccccgctgg cgcgctggct
      721 ggccgcggcc ttcgggctga cgctgctgct cgccgcgctg cgcccttcgg ccgcctactt
      781 cgggctgacg ggcagcgagc ccctgaccat cctcccgctg accctggagc cagaggcggc
      841 tgcccaggcg cactacaagg cctgcgaccg gctgaagctg gagcggaagc agcggcgcat
      901 gtgccgccgg gacccgggcg tggcagagac gctggtggag gccgtgagca tgagtgcgct
      961 cgagtgccag ttccagttcc gctttgagcg ctggaactgc acgctggagg gccgctaccg
     1021 ggccagcctg ctcaagcgag gcttcaagga gactgccttc ctctatgcca tctcctcggc
     1081 tggcctgacg cacgcactgg ccaaggcgtg cagcgcgggc cgcatggagc gctgtacctg
     1141 cgatgaggca cccgacctgg agaaccgtga ggcctggcag tgggggggct gcggagacaa
     1201 ccttaagtac agcagcaagt tcgtcaagga gttcctgggc agacggtcaa gcaaggatct
     1261 gcgagcccgt gtggacttcc acaacaacct cgtgggtgtg aaggtgatca aggctggggt
     1321 ggagaccacc tgcaagtgcc acggcgtgtc aggctcatgc acggtgcgga cctgctggcg
     1381 gcagttggcg cctttccatg aggtgggcaa gcatctgaag cacaagtatg agacggcact
     1441 caaggtgggc agcaccacca atgaagctgc cggcgaggca ggtgccatct ccccaccacg
     1501 gggccgtgcc tcgggggcag gtggcagcga cccgctgccc cgcactccag agctggtgca
     1561 cctggatgac tcgcctagct tctgcctggc tggccgcttc tccccgggca ccgctggccg
     1621 taggtgccac cgtgagaaga actgcgagag catctgctgt ggccgcggcc ataacacaca
     1681 gagccgggtg gtgacaaggc cctgccagtg ccaggtgcgt tggtgctgct atgtggagtg
     1741 caggcagtgc acgcagcgtg aggaggtcta cacctgcaag ggctgaattc tgaagggtgg
     1801 gcgcgccgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1861 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1921 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1981 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2041 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2101 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2161 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2221 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2281 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2341 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2401 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2461 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2521 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2581 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2641 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2701 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2761 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2821 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2881 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2941 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3181 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3421 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3661 ttgctggcct tttgctcaca tgtt
//
