LOCUS       sequence-118659-        6114 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 6114)
  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 6114)
  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-Sema4f-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2211..2230)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2212..2230)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2237..2254)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2237..2253)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2246..2268)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2394..2822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2481..2500)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2691..2712
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3166..3960
                     /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(3220..3239)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3830..3849
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3978..4046,4047..4838)
                     /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(4196..4215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5009..5597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5498..5517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5751..5768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5885..5906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5921..5926,5927..5944,5945..5951)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5959..5975
                     /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     5964..5986
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5983..5999
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5983..5999
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6017..6035
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6017..6034
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttcctggctt tagatagacg tctcatcgca cgtggccaga ggagccccag tgagccggat
       61 gtgggctggg ctcgggcaag aatccagcag gaagggtgga gggaagccac caaaaccact
      121 gccccgctta cccagggcca ggggcagccg ttcatcttca ggtgaaggag agggagggtc
      181 ttggctatag cttgtggtcc cagaaggtgg agcccccaga tctaagccca ccttgtctct
      241 agctagaagc tccctctgtc gccgccgctg ctgacggcga ccaatcagga ggagagtgag
      301 agatgctgct agaacaccca ggaaaaagcc aaccagccca gcccccacaa cggtgtgggc
      361 ccggtttgcg ggccccctct ggctgtccca caccaagcta taagcagcca ccacacgggc
      421 ggctccaccc tcctgacact cgcaagcata agcccccatg gcccctgggg tcaccaccac
      481 ctctagccca tcccttcgtg gggtgagcga agtcactcca ctgggctggt gccacacaca
      541 ggatgcccag gcagaactgg gggagcacgg caggaccacg tggcccaccg tagccactgg
      601 aacttcaaac actaccggat gttctccagg ttcttttgga cacaaagaag agacatctgc
      661 tgactctatg tcttgaacca tcccacggtg ctcgcctgca tgggccacac aagcatcgag
      721 ccggaagctc caggcacaca cgggatcctg ggccaggata cattctgagc agctctggag
      781 acggccacag ttgctggtgt tcacttgtgt cacctcagta tgggagccca ccagaagcca
      841 atcatggtac aatttcatgc tttcaaccgg ctgtgtttcc gggaacaagg ccagatcctc
      901 caggacactg agctgagctc cgatgcgcac agcccgatgg aggtgcccat cctctgtccc
      961 caggtagagc acatcatatt ctttccctga gaggctggtc acccggtgag ccacaactct
     1021 gagataggct gtatccgtag tgaccagcag ggggcggcca tcagctggaa acacgggcct
     1081 gtccatgagc gggtggtctc tgataaaggt gagcacacgg tctggcaggg agagtgagga
     1141 tccaaactgc tggaacttca tgttgttggt gatgcactct ccaggtctgg gctggggcac
     1201 ctcgttgtcc atgacaggca gtcccctgtt gcagtcatgt tttagctctc taaagggacc
     1261 attcagcact gcccggatgt cttggggtcg gaaggcacac acagcagaaa tggcggctcc
     1321 ttcccactgg gaggaaaaga tgccataaaa gaggggggtc cccgcgccag gctgaggtcg
     1381 aagctctgtc atatcctgca gaacccccga ggcccttcca tgctcgggcc ctggacacag
     1441 caggtcagcc tttagaaacg tcgtccatct ctgctgaagg gtcttccggc ccccaaggtc
     1501 ccccgcacac acacgggcca ctcgtgggac cttgatgcgc tcatatgagt ccaagactcg
     1561 ggaggtctcc gtgaagaaaa agaagatttc atcgtctcca tcttcatccc cccactcagc
     1621 cgggctcaag accatagctg cgacaaaggc tggggcatta agccaggatg acaaggtctc
     1681 tgttcgaatc cagtcctcag ctcgacccac agctcgggag ataatcggct ctgtccccag
     1741 gaagtttttc acagtggcgg tgtagaggac gcccccagcc attacagctg ctgaccgttg
     1801 agctggctca aaaggacatt tcccccggcc actctcaagt ctttcaacct gctggaaact
     1861 ggacacatca ataaccccgc acttcggatc aaaagcgaag gtgccacacg tgaggaggtg
     1921 agaggcattg gcaatggcga gaatctggat aaaattgtga cattcgtcct ctttcttgcc
     1981 tttcttccta cagttctgtc tgtgagtctc gggcaccatc cagtcgatcc ttcgaggctt
     2041 ttccccagag aagggcaggg ttaaagcgaa gatgctgtcc cgtgcaccga cgtaaagtgt
     2101 gtgagaagca ggatccacca ggagagcaga gtaattgtac gaagggcgaa ttctgcagat
     2161 atccatcaca ctggcggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg
     2221 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2281 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2341 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     2401 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2461 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2521 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2581 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2641 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2701 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2761 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2821 ttttaacaaa attcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
     2881 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     2941 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     3001 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     3061 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     3121 aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3181 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3241 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3301 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3361 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3421 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatcc
     3481 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3541 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3601 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3661 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3721 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3781 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3841 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3901 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3961 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4021 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4081 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4141 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4201 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4261 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4321 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4381 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4441 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4501 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4561 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4621 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4681 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4741 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4801 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4861 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4921 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4981 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5041 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5101 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5161 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5221 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5281 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5341 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5401 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5461 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5521 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5581 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5641 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5701 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5761 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5821 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     5881 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     5941 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6001 atgattacgc caagctattt aggtgacact atagaatact caagctatgc atcaagcttg
     6061 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc cctt
//
