LOCUS       sequence-166144-        3653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use to synthesize riboprobes for in situ hybridization (ISH) probe 
            for En2. Not for gene expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3653)
  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 3653)
  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-En2
FEATURES             Location/Qualifiers
     source          1..3653
                     /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(778..796)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(779..796)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(814..830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(814..830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(827..849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    838..854
                     /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(862..868,869..886,887..892))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    907..928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1045..1062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1216..1804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1296..1315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1975..2766,2767..2835))
                     /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     2598..2617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2836..2940)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3018..3473)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3155..3176)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3367..3386
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3599..3621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3613..3630
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3614..3630
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3637..3653,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3637..3653,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tgaagatgat tccaactcgc tctgccaggc cccaaggcct atctcggtga gggcttttgg
      121 gtcagggcta gctgtcccgg cagcaaagcc ttcaagcctt caggacacac tcccactgct
      181 ccccaggccc caacgctggc cacggtggcg ggagagttgg cgctggcagg tcgctacggg
      241 gctgtggcgg acaggggtca tcgacacaga cgcagacaca cggtggctgc cagcacgggc
      301 ctctgagatg gttgagaaga gaggctgcgt aggaagcagg cggagaggtt ccgggagagc
      361 tgctggcctg caaggacttt ctcagacctc agcaaagaac tgcgccctct ctccccacca
      421 gagctctcca tcaccactct ggctggcgct tcctacagtg ctgtccgagc cagcacaaag
      481 caagcggcag agcccacaga ccaaatagga gctatttatt cggttcacag tcctgtgaaa
      541 tgcagcaata aaaatctttg gacttcagca agttgtttta aatttttttt ctttcggggg
      601 aagaagtgtc caatgtccca taatggaatt taagggatta aaaaaaggga gaaaagaaaa
      661 atgaaagaaa gatctgaaac cctaaacagt cccctttaat cactagtgaa ttcgcggccg
      721 cctgcaggtc gaccatatgg gagagctccc aacgcgttgg atgcatagct tgagtattct
      781 atagtgtcac ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      841 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      901 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      961 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1021 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1081 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1141 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1201 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1261 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1321 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1381 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1441 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1501 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1561 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1621 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1681 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1741 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1801 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1861 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1921 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1981 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2041 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2101 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2161 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2221 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2281 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2341 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2401 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2461 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2521 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2581 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2641 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2701 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2761 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2821 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2881 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2941 cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga
     3001 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3061 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3121 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3181 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3241 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3301 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3361 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3421 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     3481 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3541 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3601 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact ata
//
