LOCUS       sequence-333457-        3956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a part of Chinese soft-shell turtle Wnt7b for in situ 
            hybridization probe..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3956)
  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 3956)
  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-tWnt7b
FEATURES             Location/Qualifiers
     source          1..3956
                     /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(2667..3045)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     2992..3011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3230..3334)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3360..3815)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3497..3518)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3709..3728
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3942..3956,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3956,1..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 ttgatatgca cagaaacttt cgcaagtgga
      121 ttttctacgt gtttctatgc tttggagtca tctatgtcaa attaggagct ttgtcatctg
      181 tggtggcttt aggagcaaat ataatctgca acaaaatccc aggcttggcc cctcggcaac
      241 gagcgatctg ccagagccgc cctgatgcca tcatagtgat cggagaaggg gcacagatgg
      301 gaatcaatga atgccagtat cagttccgct atggcaggtg gaactgctca gcactaggcg
      361 agaaaacagt ctttggacaa gagcttcgag taggaagcag ggaagcagcg ttcacctatg
      421 ccattactgc tgctggggtt gcacatgctg tcactgccgc ctgcagccag ggcaacctaa
      481 gcaactgtgg ctgtgaccga gagaaacagg gctactacaa tcaggaggag ggctggaaat
      541 ggggaggctg ctctgcggat atcaaatacg gcattgaatt ctccaggaag tttgttgatg
      601 cgcgagaaat caaaaagaat gcccggaggc tgatgaactt gcacaacaat gaggctggaa
      661 gaaaggttct tgaagagaga atgaaattgg aatgcaagtg tcatggcgtt tcaggttcct
      721 gtacaactaa gacctgctgg accacacttc caaaattcag agagattgga tatattctga
      781 aagaaaagta caatgctgca gtacaagtgg aggtggtacg ggccagtcga ctacgacagc
      841 caaccttcct gaagataaag cagatgagga gttatcagaa accaatggaa acagatctgg
      901 tgtatatcga aaagtctccc aactactgtg aggaagatgc ttccacgggg agcgtgggta
      961 cccagggacg actctgtaac cgtacgtctc ctaatgcaga cgggtgtgac atgatgtgtt
     1021 gtggaagggg ttacaacaca caccagtaca caaaagtgtg gcagtgtaat tgcaaattcc
     1081 actggtgctg ctttgatcga 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 gagccacgct
     2521 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2581 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2641 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2701 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2761 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2821 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2881 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2941 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3001 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3061 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     3361 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3421 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3481 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3541 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3601 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3661 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3721 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3781 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     3841 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3901 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
