LOCUS       sequence-146370-        5072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use to synthesize riboprobes for in situ hybridization (ISH) probe 
            for Ascl1 (also known as Mash1). Not for gene expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5072)
  AUTHORS   Hirota J, Mombaerts P
  TITLE     The LIM-homeodomain protein Lhx2 is required for complete 
            development of mouse olfactory sensory neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8751-5. Epub 2004 Jun 
            1.
  PUBMED    15173589
REFERENCE   2  (bases 1 to 5072)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2004-06-8"; volume: "101(23)"; pages: 
            "8751-5. Epub 2004 Jun 1"
COMMENT     Sequence Label: pISH-Ascl1
FEATURES             Location/Qualifiers
     source          1..5072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2194..2212)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2195..2212)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2230..2246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2230..2246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2243..2265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2254..2270
                     /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(2278..2284,2285..2302,2303..2308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2323..2344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2461..2478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2632..3220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2712..2731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3391..4182,4183..4251))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4252..4356)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4437..4892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4574..4595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4786..4805
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5018..5040
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5032..5049
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5033..5049
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5056..5072,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5056..5072,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc tctagactcg aggaattcga ggatccgggt
       61 accatgtgcc ctttgatcgt gcttcgcagc cccgcttcct tcaagggctc tgcgcaccct
      121 gcgatcccca actcgttctc ccccgcgaca gtttggcccg gcatggagag ctctggcaag
      181 atggagagtg gagccggcca gcagccgcag cccccgcagc ccttcctgcc tcccgcagcc
      241 tgcttctttg cgaccgcggc ggcggcggca gcggcggcgg ccgcggcagc tcagagcgcg
      301 cagcagcaac agccgcaggc gccgccgcag caggcgccgc agctgagccc ggtggccgac
      361 agccagccct cagggggcgg tcacaagtca gcggccaagc aggtcaagcg ccagcgctcg
      421 tcctctccgg aactgatgcg ctgcaaacgc cggctcaact tcagcggctt cggctacagc
      481 ctgccacagc agcagccggc cgccgtggcg cgccgcaacg agcgcgagcg caaccgggtc
      541 aagttggtca acctgggctt tgccaccctc cgggagcatg tccccaacgg cgcggccaac
      601 aagaagatga gcaaggtgga gacgctgcgc tcggcggtcg agtacatccg cgcgctgcag
      661 cagctgctgg acgagcacga cgcggtgagc gctgcctttc aggcgggcgt cctgtcgccc
      721 accatctccc ccaactactc caacgacttg aactctatgg cgggttctcc ggtctcgtcc
      781 tactcctccg acgagggatc ctacgaccct cttagcccag aggaacaaga gctgctggac
      841 tttaccaact ggttctgagg acctgccagg ctctcctggg aatggacttt ggaagcagga
      901 tggcagcaga tcctgcatct ttagtgtttc tcgccaacga cgtcaaatgg ggaggcagaa
      961 aaacaagggg aaaaaagaag aagaaatgaa acaaacaaac cagacagcca acctacaggg
     1021 gcaccttcac taagatgcaa tgttctcagc aaacaggggt gggctccaac agtgtctctg
     1081 cattccaaca tcatttccag acacgagaag agtgactggt gtctgaacct aagcccgaat
     1141 cacagatggg ttcctttcct ggagcaagag cgtcacacac acacacacac acacagacag
     1201 acactatatt aactcccaac cactaacagg cagggctgga agcgcgcatg tgcaagtgcc
     1261 ttcacctccc actctctgtc agagctgtct tagccccctg aaactgggtt gatgtctttc
     1321 ctcagtcacc cccattccag cgatctatgg acatttgcct ccattgaagc aacgtcagtt
     1381 ctcggacagc ctttccctct cctggtggcc tcctccccaa accccacatc gccctcccac
     1441 ggtctttgct tctgttttct tcatagaatg cttccaatct ttgtgaattt ttttattata
     1501 agaaaaaaaa tctatttgta tctatcctaa ccagtttggg gatatattaa gatatttttg
     1561 tacataagaa aaagagagag aaaaaattta tagaagtttt gtacaaatgg tttaaaaatg
     1621 tgtatatctt gatactttaa catgtaatga gattacctct gcgtacttta gatatgtagt
     1681 tcatcttaca actgccatcc ccacccccat ccccagtgtg gttttggaaa gaactctcct
     1741 cataggtgag atctaaatgc caccagaatg acttcagcac caatgtgtct tacttcacag
     1801 aaacgtggtt aatgtattaa tgatgttatt aaaaaaaaac tgttcaagaa gaacaaaagt
     1861 ttatgcagct actgtccaaa cgcaaagtgg cagcccgttg gttcccatag gttgcctttt
     1921 ggaagtttct accattgcct tccccctcac acccacccct tactgtttta ttacaaactt
     1981 acaaaaaaaa gtgtataacc ctgttttata caaactagtt ttgtaataaa actttttcct
     2041 tttttatata aaaagaagag agagagagaa tgaaaaaaaa aaaaaaaaaa aaaaaccatg
     2101 gtacccggat cctcgaattc ggtaccccgg gttcgaaatc gataagcttg gatccggaga
     2161 gctcccaacg cgttggatgc atagcttgag tattctatag tgtcacctaa atagcttggc
     2221 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2281 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2341 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttcgatag
     2641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2761 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2821 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3061 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3181 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3241 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3301 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3661 aagtagttcg ccagttaata gtttgcgcaa cgttgttggc attgctacag gcatcgtggt
     3721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3961 ctgagaatac cgcgcccggc gaccgagttg ctcttgcccg gcgtcaatac gggataatag
     4021 tgtatgacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4381 tgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgaaat
     4441 tgtaaacgtt aatattttgt taaaattcgc gttaaatatt tgttaaatca gctcattttt
     4501 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4561 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4621 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     4681 aagttttttg cggtcgaggt gccgtaaagc tctaaatcgg aaccctaaag ggagcccccg
     4741 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4801 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4861 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     4921 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4981 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5041 acggccagtg aattgtaata cgactcacta ta
//
