LOCUS       sequence-118664-        5933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains 2kb cDNA fragment for probe synthesis for mRNA in situ 
            hybridization, species: mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5933)
  AUTHORS   Maier V, Jolicoeur C, Rayburn H, Takegahara N, Kumanogoh A, Kikutani
            H, Tessier-Lavigne M, Wurst W, Friedel RH
  TITLE     Semaphorin 4C and 4G are ligands of Plexin-B2 required in cerebellar
            development.
  JOURNAL   Mol Cell Neurosci. 2011 Feb;46(2):419-31. doi: 
            10.1016/j.mcn.2010.11.005. Epub 2010 Nov 29.
  PUBMED    21122816
REFERENCE   2  (bases 1 to 5933)
  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.mcn.2010.11.005"; journalName: "Mol Cell Neurosci"; date:
            "2011-02"; volume: "46"; issue: "2"; pages: "419-31"
COMMENT     Sequence Label: pCRII-Plxnb2-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..5933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2030..2049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2031..2049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2056..2073)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2056..2072)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2065..2087)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2213..2641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2300..2319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2510..2531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2985..3779
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3039..3058)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3649..3668
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3797..3865,3866..4657)
                     /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     complement(4015..4034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4828..5416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5317..5336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5570..5587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5704..5725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5740..5745,5746..5763,5764..5770)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5778..5794
                     /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)."
     primer_bind     5783..5805
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5802..5818
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5802..5818
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5836..5854
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5836..5853
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gtaggttgtg tgccgtgtct cgcttctgcc gccgcttggg tggaggctgc acctctgggg
       61 gctcgcagta caggtccgtc tcggtcagcg tcttcataat gcaacgctca gcacccacaa
      121 atgcctctgc ctcctccaga gtcatggctt tgttcagatt ggttccccgg gcgtggatga
      181 gcttgttgac ctgctttttg acgccacccg tgaagttctc aaaggtgggg tctgccacat
      241 actcaaagac tccggcttcg gtcctgagcg gggcacagtg accgtccatc cgtatcagca
      301 cggtgaggtt gtaagcctcg ggctcttcgg ggacagcagg agacaagaag acgaccttgg
      361 tgtcgttgta gaacacgtac tccatgccct cgaccgtcac acgctccagg gctccagcct
      421 cccgccgccg ccgcctccag gaccgcaagg gctcagcgat gacaaccatg gcaaacttct
      481 ggatgaggct gaagccctgg ccagtaacgt tgatgctccg gccaccactg acaaagcttc
      541 tcaatggctc aaaggctcgt atcatggggt tctcgcagta ggtgaaagag atgccggggc
      601 tgggcactct ggagccccca tagtagattt ctagtgtcac ctggcctgga gccaactgtc
      661 gacctgtgac acactgtagc tgtgctccaa actttgtcac ttcacaaggg acgtcattga
      721 gtgtcacccg cacatcctcc ttggagcctg tgtccaggtg agtgccattg atggtcaatg
      781 tggtgccacc tgcctgtggc ccctgtcgtg gctccacact gagaggctgg ggttgttgat
      841 aagtgaactg gacgtgtggc ggtgaatggc cgagttttcc attaacatcc acctcaatgc
      901 ctcctgtgaa gggcatctcc gaagcctcaa ttgcacacgc aatccgggtg gatacggagt
      961 accctcttgg ttcaaaggca cagttctggc cagccacagt tatcttcttg acgtcatctg
     1021 ctttgacacc cagattggac ccatggatag tgaccaggat gcccccaccc agcgggcccg
     1081 tctcaggctg gatcctagtg ataactggtg gtgggcactc agaagtgaca ttgctgcaca
     1141 gagcttcgta cacgcaccgg ttctgcccac ggcaccacac acacctgtag gcaggatcgg
     1201 cagccagaca caggctacag tcactgcggc caaaggagca gttgtagaga gtcacttgta
     1261 gcttgctgtc aatgttcttg ccaaaggact taacatacag gtgaagaggc agtgtctcat
     1321 taccatcatg ggatagcttt ggggtcctaa aagaggaggt gtctgattca tgcatggtca
     1381 cagtctcttc aaaattcagc agctcactgc ccacatacag ggaagaaacc ttcacagtct
     1441 ccaagttctt gccctggaag gtcacctctg tctcatggtt catggggatg accaaagggt
     1501 caggggccag gaactggggg cagttgtcct ccatgtgggc acgtatgatt ccctcctctg
     1561 ggttgggcga agcctcccga cactcgtagt actgcaggtc ccattggcaa gtccagcggt
     1621 tgctagcaca agagatgcac ggcaggttct ccaccaggct catggcctca cggcagtcat
     1681 agaagggata ctggtgggag gtgaggaaga cactgccgct ttttaagagg agctggatgc
     1741 tcacgtcaac atggtcttgg cctggcggtg tactagggat gctgcttggg gaattacaga
     1801 tgacggtatc gtcctctacc cgggcagggt ggggtggtga gtcaccaaag aggcacagta
     1861 actcatcgtc ctcagtcagg gtgggcaggg ggctgacaga caggcgtacc tctccttggg
     1921 cccgccggct catgttctgt ggaaaggcat cagtgatggc aagggcgaat tctgcagata
     1981 tccatcacac tggcggccgc tcgagcatgc atctagaggg cccaattcgc cctatagtga
     2041 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2101 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2161 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2221 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2281 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2341 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2401 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2461 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2521 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2581 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2641 tttaacaaaa ttcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     2701 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     2761 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     2821 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     2881 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     2941 ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3001 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3061 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3121 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3181 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3241 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatccc
     3301 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3361 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3421 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3481 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3541 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3601 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3661 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3721 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
     3781 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3841 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3901 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3961 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4021 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4081 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4141 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4201 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4261 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4321 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4381 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4441 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4501 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4561 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4621 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4681 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4741 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4801 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4861 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4921 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5221 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5281 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5341 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5401 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5461 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5521 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5581 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5641 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5701 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5761 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5821 tgattacgcc aagctattta ggtgacacta tagaatactc aagctatgca tcaagcttgg
     5881 taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcgcc ctt
//
