LOCUS       sequence-166182-        3527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use to synthesize riboprobes for in situ hybridization (ISH) probe 
            for Pitx1. Not for gene expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3527)
  AUTHORS   Parrilla M, Chang I, Degl'Innocenti A, Omura M
  TITLE     Expression of homeobox genes in the mouse olfactory epithelium.
  JOURNAL   J Comp Neurol. 2016 Oct 1;524(14):2713-39. doi: 10.1002/cne.24051. 
            Epub 2016 Jun 19.
  PUBMED    27243442
REFERENCE   2  (bases 1 to 3527)
  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: "10.1002/cne.24051";
            journalName: "J Comp Neurol"; date: "2016-10-1- 1"; volume: "524"; 
            issue: "14"; pages: "2713-39"
COMMENT     Sequence Label: pISH-Pitx1
FEATURES             Location/Qualifiers
     source          1..3527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      33..42
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(652..670)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(653..670)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(688..704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(688..704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(701..723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    712..728
                     /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(736..742,743..760,761..766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    781..802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(919..936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1090..1678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1170..1189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1849..2640,2641..2709))
                     /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     2472..2491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2710..2814)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2892..3347)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3029..3050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3241..3260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3473..3495
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3487..3504
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3488..3504
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3511..3527,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3511..3527,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tcccccagct ccatctcttc catgaccatg ccgtccagca tgggcccggg cgccgtgccc
      121 ggcatgccca actcgggcct caacaacatc aacaacctca ccggctcctc gctcaactcg
      181 gccatgtcgc cgggcgcctg cccctacggc accccagcct cgccctacag cgtctaccgg
      241 gacacgtgca actcgagcct tgccagcctg cggctcaagt ccaagcagca ctcgtcgttt
      301 ggctatggcg gtctgcaggg tccggcgtcg gggctcaacg cttgccagta caacagctga
      361 ccgccccgcc tggtccccgg ggccgccggc cagcaaggct cacagaaggg agcgcgcggg
      421 agcacgcggg gcgctcgcag ctcctcggac ctcacctctg cgcagcccct tcctccccac
      481 cccgcctcca ggttggtttt gtgtttgctt tttccagacc cccgctctgt cccccaatga
      541 atgaaaagaa aagaaagcaa gccaagggag gaatcactag tgaattcgcg gccgcctgca
      601 ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta ttctatagtg
      661 tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2821 tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2881 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2941 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3001 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3061 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     3361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     3421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     3481 cgacgttgta aaacgacggc cagtgaattg taatacgact cactata
//
