LOCUS       sequence-273180-        3541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template to synthesize riboprobes for in situ hybridization (ISH) 
            for Olfr449.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3541)
  AUTHORS   Zapiec B, Mombaerts P
  TITLE     The Zonal Organization of Odorant Receptor Gene Choice in the Main 
            Olfactory Epithelium of the Mouse.
  JOURNAL   Cell Rep. 2020 Mar 24;30(12):4220-4234.e5. doi: 
            10.1016/j.celrep.2020.02.110.
  PUBMED    32209480
REFERENCE   2  (bases 1 to 3541)
  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.celrep.2020.02"; journalName: "Cell Rep"; date: 
            "2020-03-24- 24"; volume: "30"; issue: "12"; pages: "4220-4234.e5"
COMMENT     Sequence Label: pISH-Olfr449
FEATURES             Location/Qualifiers
     source          1..3541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..125
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     117..134
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     118..134
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     141..160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        141..159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      190..199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      194..203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(823..841)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(824..841)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(859..875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(859..875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(872..894)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    883..899
                     /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(907..913,914..931,932..937))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    952..973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1090..1107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1261..1849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1341..1360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2020..2811,2812..2880))
                     /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     2643..2662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2881..2985)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3063..3518)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3200..3221)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3412..3431
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
       61 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      121 aaacgacggc cagtgaattg taatacgact cactataggg cgaattgggc ccgacgtcgc
      181 atgctcccgg ccgccatggc ggccgcggga attcgatttc ttccttgcca acctgtcttt
      241 cttggagacc tggtacatct ctgttaccgt gcccaagctg ctgttcagct tctggtccat
      301 gagtaacagc atttctttca ctcactgtat gatacagctt tacttcttca ttgcacttat
      361 gtgtacggaa tgcgttctcc tagctgcaat ggcctatgat cgttatgtgg ctatctgccg
      421 cccattacac taccccacca tcatgagcca tggactctgt ttccgcctag ctcttggttc
      481 ctgggtcatt ggctttggca tctctttggc aaagatctac ttcatctccc gcctcagctt
      541 ctgtggcccc aatgtcatca accacttctt ttgtgatatc tctcccgttc ttaatctctc
      601 ctgcacagac atgtccattg cagagctggt ggactttgtc ttggctctgg tcatcttcct
      661 ctttccgctg tcaatcactg ttttatcata cggatgcatt ctggccactg tgttacgcat
      721 gccaacagga aagcagaaag ccaatcacta gtgaattcgc ggccgcctgc aggtcgacca
      781 tatgggagag ctcccaacgc gttggatgca tagcttgagt attctatagt gtcacctaaa
      841 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
      901 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
      961 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1021 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1081 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1141 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1201 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1261 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1321 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1381 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1441 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1501 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1561 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1621 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1681 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1741 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1801 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1861 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1921 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1981 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2041 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2101 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2161 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2221 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2281 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2341 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2401 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2461 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2521 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2581 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2641 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2701 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     2761 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     2821 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     2881 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2941 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3001 agtgccacct gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3061 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3121 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3181 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3241 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3301 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3361 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3421 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3481 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc
     3541 a
//
