LOCUS       sequence-166202-        3629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use to synthesize riboprobes for in situ hybridization (ISH) probe 
            for Six4. Not for gene expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3629)
  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 3629)
  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-Six4
FEATURES             Location/Qualifiers
     source          1..3629
                     /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(754..772)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(755..772)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(790..806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(790..806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(803..825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    814..830
                     /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(838..844,845..862,863..868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    883..904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1021..1038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1192..1780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1272..1291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1951..2742,2743..2811))
                     /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     2574..2593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2812..2916)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2994..3449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3131..3152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3343..3362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3575..3597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3589..3606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3590..3606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3613..3629,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3613..3629,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tcctttatgc tgttagtcca gggcatctct gatttcagaa agaaggcgac tttttctaag
      121 gtgacttctc agttgctaag cacaaagata gtaccactcc ttggttttgt taaggtggtg
      181 gaagggtttc taactaacac actcatcagc tgagcctgca gagtaacggt cactggggac
      241 ttaacatttg ttgttacaca agggaacttt cagtcaagat taagagccct atgaagaatt
      301 ctgtgcccat cattttgatc aagaacctaa atcccatttt tgtgaaattg gtgaatgtcc
      361 ctcaattgct gaaatatgca atttgggcca gaggttgttg tttgtttgtt ttggatgttc
      421 ttgttttatt ttcatgaagc cagttgcttc agtgtggttt tccactctga cactacttac
      481 acagcttggc tgcccccgtc tcccagttat ttactgatca tattaatgtg attagcatcc
      541 tggtcttgta gtatagttta catttttaaa aaatgttatt ttttactaga ttgctgcaga
      601 tatcaagagg acattatttt aaaatacctg tctggtaagg gaagtgggaa agtcaaaacg
      661 ttctataagg cggaatcact agtgaattcg cggccgcctg caggtcgacc atatgggaga
      721 gctcccaacg cgttggatgc atagcttgag tattctatag tgtcacctaa atagcttggc
      781 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      841 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      901 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      961 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2941 tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     3001 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3061 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3121 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3181 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3241 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3301 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3361 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3421 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     3481 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     3541 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     3601 gccagtgaat tgtaatacga ctcactata
//
