LOCUS       sequence-333460-        3958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a part of Chinese soft-shell turtle Wnt5a for in situ 
            hybridization probe..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3958)
  AUTHORS   Nomura T, Ohtaka-Maruyama C, Kiyonari H, Gotoh H, Ono K
  TITLE     Changes in Wnt-Dependent Neuronal Morphology Underlie the Anatomical
            Diversification of Neocortical Homologs in Amniotes.
  JOURNAL   Cell Rep. 2020 May 5;31(5):107592. doi: 
            10.1016/j.celrep.2020.107592.
  PUBMED    32375034
REFERENCE   2  (bases 1 to 3958)
  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: "Cell Rep.";
            date: "2020-05-5"; volume: "31(5):107592. doi"; pages: " 
            10.1016/j.celrep.2020.107592"
COMMENT     Sequence Label: pBS-tWnt5a
FEATURES             Location/Qualifiers
     source          1..3958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     67..83
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     68..84
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1118..1134)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1118..1134)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1171..1189)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1210..1226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1210..1226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1223..1245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1234..1250
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1258..1264,1265..1282,1283..1288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1441..1458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1612..2200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1692..1711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2371..3162,3163..3231))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2994..3013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3232..3336)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3362..3817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3711..3730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3944..3958,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3958,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggtaccgggc
       61 cccccctcga ggtcgacggt atcgataagc ttgatatggc ttccaagtac ctcatagtgg
      121 ttctggccat tttctcctca ttcgcccagg ttgtaataga agccagctct tggtggtctt
      181 taggaatgaa tcatatgaat cctatgaacc ctgttcagat gtcagaggta tatataatag
      241 gagcacagcc actatgtagt caattagcag gactttccca aggacaaaag aaactctgcc
      301 acttgtatca ggaccacatg cagtatattg gagagggggc aaagacgggc attaaggaat
      361 gccagtatca gttcagacac agaagatgga attgcagcac tgtggacaac aactctgttt
      421 ttggcagagt catgcagata ggcagtaggg agactgcctt tacttatgca gtcagtgctg
      481 caggggtggt taatgcaatg agccgtgcct gccgagaggg ggagctctcc tcctgtggct
      541 gcagccgagc agccaggccc aaagatttac cccgggactg gctgtggggt ggctgtgggg
      601 ataacatcga atacggctat cgcttcgcca aggaatttgt ggatgctcgg gagcgtgaga
      661 gaatttatca gaaaggctct tacgagagtg ctagaatttt gatgaacctg cataacaacg
      721 aggctggaag acgaactgtg tataacttgg cggatgtggc ttgtaagtgt catggggtgt
      781 ctggctcctg cagcctgaag acctgttggc tgcagctggc cgatttccgc aaggtgggcg
      841 atgccctgaa ggagaaatat gatagcgctg ctgccatgaa actcaatagc cggggcaagc
      901 tggtgcaagt gaacagccgt ttcaacacgc ccaccatcca tgacctggtc tacattgacc
      961 ctagccccga ctattgtgtc cgaaatgaga gtactggctc cctgggcacc cagggccgcc
     1021 tctgcaataa gacttcagag ggtatggatg gctgcgaact catgtgctgt ggtcgggggt
     1081 acgaccagtt caagaatcga attcctgcag cccgggggat ccactagttc tagagcggcc
     1141 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattt cgagcttggc
     1201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3361 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3421 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3481 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3541 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3601 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3661 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3721 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3781 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     3841 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3901 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgtt
//
