LOCUS       sequence-118666-        5953 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 5953)
  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 5953)
  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-Plxnb3-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..5953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2050..2069)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2051..2069)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2076..2093)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2076..2092)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2085..2107)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2233..2661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2320..2339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2530..2551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3005..3799
                     /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(3059..3078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3669..3688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3817..3885,3886..4677)
                     /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(4035..4054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4848..5436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5337..5356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5590..5607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5724..5745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5760..5765,5766..5783,5784..5790)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5798..5814
                     /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     5803..5825
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5822..5838
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5822..5838
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5856..5874
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5856..5873
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ggtggacatc atggactctg cttcgtactg gacagggcct agagccaggc gcaggttgcc
       61 catctgcacc acaaactgcg gcagggtgcc tgaaccattg gtgggttgag gggcttgggg
      121 tggtggctca cagtacaagt gagtaagcgt cagtgtcttc accaggcact ctccatctcc
      181 aatatgcacc tgcacctcct ccttgctgat ccccagattg aggccctcgc cctccacatc
      241 taggacatgg cctggcttga ggtggtaggg gtgagtaatc ccctcatgac taagtggagc
      301 caggcgaggg ttaggctggt ataggaatcc ctggccccca ctagcactgg caaagtctac
      361 ttgcatgttg tctagggcaa agaagactcg ctttgggagg gccccatctg gcactgctgg
      421 actgtggcac aggaggaggc tggatgagtt gacagagcag agcgtggaac actgtagtaa
      481 gccactctca agatggatac aggcctgagg atctgcagca ggggcaccac agctcctcct
      541 tgggttcgca tcctgggcct gaacccccaa ggcctttact ttgggctcat cctccagcca
      601 cacagacagc aagggctgcc acacaacgtc taagcctgta cccctgactc gaatcactcg
      661 tccacctccc cggaagctga cactaggctc tgcttccacc agttggggat tggctgtgta
      721 tcgaaaaggg gtggtgagca gtttgcgttc aacatgccca aagactatga ggaccactgc
      781 ttctcctggc tcagtctggg gcatggtgtg gcatatgatg gcttcaggac acacaggctc
      841 ctggataggg caaggttggt cacccacaaa gacactgatg ttgcctcctg tctggagata
      901 ctgcccatgg atggtgagct gggtgccccc tgcctggggg ccccactgag gattcaggct
      961 cagcaggaca gggtcctggt aggtgaagtt ctgggttgag atgccaggag gacgactctt
     1021 aatggccact tggactggcc cagcagtgcc attgggagca ggagatgtca cacatacaat
     1081 ccgagcagag atgcggtaga gagaagggtc agggttgcag ggctggccag ccacagttac
     1141 agcatttcgc acatcattaa aggcctggcc caggttggaa cccaggatgg tgatggccaa
     1201 gccaccttct gggggaccag tcaggggttc aatcacatca atgctgggta tagggcatag
     1261 ctgttccaca gcacctggtg ggcacaaagg cccatagcgg caggcaggct ggccatcacc
     1321 acaccacaaa cagctcaggc tcccattggc cgcctggcag tggctgcagt cagggtgacc
     1381 cgtggcacag tcatatagag tcacatgaag atcaccagca ttgtccagcc gctggatctc
     1441 acccctggtt acatagatgg gcactgggag ctcccactga gacatagagg ggtagaactg
     1501 ctgggcctgg cagtgaatga ggccagagtc cctggatgtc tcctctaagg aggcaggtag
     1561 cttttgcagt tttccaggca gctctagcca gcagtggtac aaggcaggca gaccttggaa
     1621 atagtgaagg ttctgaatgt gcagggtcac atggctctcc caacccacag gcaccaactg
     1681 gggacttgct agaccctcaa cctgggggca agcctctggg ccacgaacca ggatgtccac
     1741 ctcctgggca ctgtagacag ccttttcacc ctctgggcag tgctctccat atatacagtg
     1801 gctactctgg gggcaccagt ggcaccgcca caggctgctc acacaagcgc ggcacggggc
     1861 agccacttcc aaggcctgaa cagcactgca gtcatagaag gagaatgtgg tggcggtcaa
     1921 taccacatcc tcaaacatta gggccagggg caaggtgaca tgatctgagc ctggaggatt
     1981 aagggcgaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
     2041 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     2101 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2161 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2221 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2281 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2341 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2401 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2461 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2521 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2581 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2641 aacaaaaatt taacgcgaat tttaacaaaa ttcagggcgc aagggctgct aaaggaagcg
     2701 gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     2761 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     2821 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     2881 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     2941 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
     3001 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3061 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3181 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3301 gcaggatctc ctgtcatccc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3781 tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3841 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3901 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3961 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4021 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4081 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4141 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4201 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4261 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4321 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4381 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4441 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4501 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4561 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4621 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4681 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4981 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5521 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5581 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5641 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5701 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5761 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5821 acaggaaaca gctatgacca tgattacgcc aagctattta ggtgacacta tagaatactc
     5881 aagctatgca tcaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct
     5941 ggaattcgcc ctt
//
