LOCUS       sequence-118660-        6006 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 6006)
  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 6006)
  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-Sema4g-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..6006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2103..2122)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2104..2122)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2129..2146)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2129..2145)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2138..2160)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2286..2714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2373..2392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2583..2604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3058..3852
                     /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(3112..3131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3722..3741
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3870..3938,3939..4730)
                     /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(4088..4107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4901..5489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5390..5409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5643..5660
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5777..5798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5813..5818,5819..5836,5837..5843)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5851..5867
                     /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     5856..5878
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5875..5891
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5875..5891
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5909..5927
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5909..5926
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gctaccacat agaacatcct catgtcatgt gccagctgtg ccccaggtgt ggcaggggct
       61 tgtggagctg gagcaggggt ggctggccgg actgtgaggc tgtaggacgc cagcagcata
      121 cggaggccat tctcctcggc atagcagcca tagttgccac tgtgctccag ttgtgtgtca
      181 gttaccagca gcccatccac tcccacgcgg tagccatcct gcccatcgct caggctcttg
      241 ctcccattga gtagccacaa agcccgggcc aggttagacg gctggtcaca gggcaggagg
      301 acatcatcac cccggagcac agatcgggtc ttgagtggtg gtggtggtcc tgcatccctg
      361 ctgccctcac agcctcgatt tcctctctct atgtcctgta tcaattctgt cctgttggct
      421 acggtggtgg ccaccatgca ggcatggatg ctggagtccc agccacagta ggggtcccgt
      481 gccaggatgc agtcatagca ggactggtag cgtgagcagc tggacagagg aaactggagg
      541 actccacttg cagcccccac gtagaggctg tgctgcattg gtgagataac tagattgtcc
      601 acagactggg gctccctgaa cacttgtatc tcttcaatta tgtgcatccc agagcccagg
      661 actacggcct tgtggatcca gccatcagct gtgcccagaa agagcaggtc ataggtgggt
      721 ccagcaggtg tgctgacgtg tgttcctgtg agatgtgtgt agcgtacatt gcgcttcagc
      781 agcaagggcc gcccacgtgt gggcaccaca ggccgagcca tcaggggatg caacttcaca
      841 aagtccagaa ccagggatgg caagtcttgg gatgagttgt agcctcggct gcgcaatgaa
      901 tctgtgatgc acgagccagg ccggggttca ggcaccccac cctcatagcg gccccagcgc
      961 cgggcaccat cctggtactc cataaaaggg cctgtgaaga cagcctgaat ctcagccagg
     1021 tcatagcggc agatggctga agcctccagc gtcttccact gtgttgtcag tgtgaagacg
     1081 gcatagaaat gtgtgtggga tgatgtatca gcattcaggc tgcagacccc acgtagtgtc
     1141 tcatactgtg ggatgtggca gatgagacga gccttcagga aggatgtcca cttcttctgt
     1201 aggatcttct tccctcccag atctcccttg cacacacggg ctactctagc cacacggtgg
     1261 ctgctgcgac tctgtgtgaa gctgctggag ccttcctcac gccccatgaa gaagaaataa
     1321 atcttgtcat catcacccac tgcgcttgtc ttgctctccc gcaccagcac agagaacaca
     1381 aactcagcat cattgagcca gtgcatgggt gcctcctcag ttctcagaga gtgtgggtgg
     1441 cggctccggc ggatatcagg aatgctccgg aattcatacc gggtggctgt gtagaggcct
     1501 ccatcaatga taaggcctgt gaagccccgg gccgggtcat aaggacattt ctcctttccc
     1561 tcctcgaaac tagtaggcaa gatgaaggtt tcagcatcaa tagctgcaca gaggggctgg
     1621 aaggcgtggg tcccacatgc atagaagtgg gtggcattga gccgctgtag aaatcgcacg
     1681 tggttgaaac actcggtctg gttgtttttc cctttctgat gacacttgct ttgcatctct
     1741 ggggaggcct cccagtggat ctctttgtga gtcctatctc ttatgtcacg ggcactgaga
     1801 gagaacaggg cgcctcgagc ccccaccagc agcctctcag aagcctcctc cagcagcagt
     1861 gttgaatagt tctgggtttg gcccttgaag tgccgaatct gggagagctc ttcataggag
     1921 atggtcagcc gtggggtggc atctagctct cgggatggtc tccgcagtga gggtccaggc
     1981 actgctgtta ctgtgagaaa gctgaagagg aggggccaga gcctccccca cataagggcg
     2041 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga gggcccaatt
     2101 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     2161 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2221 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2281 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2341 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2581 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2641 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2701 atttaacgcg aattttaaca aaattcaggg cgcaagggct gctaaaggaa gcggaacacg
     2761 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     2821 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
     2881 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
     2941 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
     3001 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
     3061 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3121 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3181 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3241 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3301 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3361 ctcctgtcat cccaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3421 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3481 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3541 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3601 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3661 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3721 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3781 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3841 gagttcttct gaattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3901 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3961 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4021 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4081 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4141 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4201 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4261 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4321 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4381 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4441 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4501 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4561 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4621 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4681 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4741 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4801 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5041 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5521 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5581 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5641 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5701 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5761 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5821 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5881 acagctatga ccatgattac gccaagctat ttaggtgaca ctatagaata ctcaagctat
     5941 gcatcaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc
     6001 gccctt
//
