LOCUS       sequence-210064-        3672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Preparation of RNA probe for ISH of mouse Vmn2r65 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3672)
  AUTHORS   Akiyoshi S, Ishii T, Bai Z, Mombaerts P
  TITLE     Subpopulations of vomeronasal sensory neurons with coordinated 
            coexpression of type 2 vomeronasal receptor genes are differentially
            dependent on Vmn2r1.
  JOURNAL   Eur J Neurosci. 2018 Apr;47(7):887-900. doi: 10.1111/ejn.13875. Epub
            2018 Mar 25.
  PUBMED    29465786
REFERENCE   2  (bases 1 to 3672)
  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: "10.1111/ejn.13875";
            journalName: "Eur J Neurosci"; date: "2018-04"; volume: "47"; issue:
            "7"; pages: "887-900"
COMMENT     Sequence Label: pISH-Vmn2r65
FEATURES             Location/Qualifiers
     source          1..3672
                     /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(797..815)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(798..815)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(833..849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(833..849)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(846..868)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    857..873
                     /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(881..887,888..905,906..911))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    926..947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1064..1081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1235..1823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1315..1334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1994..2785,2786..2854))
                     /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     2617..2636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2855..2959)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3037..3492)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3174..3195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3386..3405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3618..3640
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3632..3649
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3633..3649
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3656..3672,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3656..3672,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 ttccccattc taagattcct ggttttaaga aattcatcca gacagtgcac ccttccaact
      121 acagtaagga tacttccctt gctaggttgt ggtggctgta tttcaactgt tcattgccat
      181 ctcattgtaa gacactgaaa aattgttcaa ccaaaatcct actagaatgg ttatccaggc
      241 accagtttga agtgtcaatg agcgagccaa gttacaacct atacaatgct gtctatgctg
      301 tagcctatgc actccatgaa atgcttattc aacatacaga acattggcag aaggatattg
      361 gaaaagaact ggaattttac tcctggaagg tggccccttt cctggagaac aacaaattta
      421 taattccttc tgaatatcaa tacagaagag ggaaactgga tacagaatat gacattctct
      481 atgctatgga tcttctacca gctcttggac ttaatgtgaa aataggaaag ttttcccaac
      541 attttctaca ttttcagcaa ttatatatag cggaagacat gatagattgg accatagaca
      601 ttagacagac attacaatca gtatgcagtg tgccctgtag tgtaggattc cgaaaattca
      661 ttcagaaagg aaaggctgtc tgctgttttg attgtacccc atgtccagag aatgaaaatc
      721 actagtgaat tcgcggccgc ctgcaggtcg accatatggg agagctccca acgcgttgga
      781 tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc
      841 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      901 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      961 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1021 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1081 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1861 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1921 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1981 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2041 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2101 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2161 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2221 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2281 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2341 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2401 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2461 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2521 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2581 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2641 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2701 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2761 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2821 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2881 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2941 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgatgcg gtgtgaaata
     3001 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     3061 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3121 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3181 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3241 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3301 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3361 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3421 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3481 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     3541 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     3601 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata
     3661 cgactcacta ta
//
