LOCUS       sequence-118657-        5910 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 5910)
  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 5910)
  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-Sema4c-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..5910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    272..293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(308..313,314..331,332..338)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    346..362
                     /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     351..373
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     370..386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     370..386
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        404..422
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     404..421
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2508..2527)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2509..2527)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2534..2551)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2534..2550)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2543..2565)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2691..3119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2778..2797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2988..3009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3463..4257
                     /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(3517..3536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4127..4146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4275..4343,4344..5135)
                     /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(4493..4512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5306..5894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5795..5814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
       61 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
      121 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
      181 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      241 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      301 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      361 aatttcacac aggaaacagc tatgaccatg attacgccaa gctatttagg tgacactata
      421 gaatactcaa gctatgcatc aagcttggta ccgagctcgg atccactagt aacggccgcc
      481 agtgtgctgg aattcgccct tacagcaaca aggtagcttt ctgcagccag tcttgtcccc
      541 tgctcctctg aatagcaacg atagggtcca gagtgacggg actgtgcggc catcaccacc
      601 agcgcctgga gtcccgtgtc ataaagaaag gagccaggtt gttctgcagg caggtcctgg
      661 cttccgaagg tccagtgggc atgggccaaa ttggacgaga ggtggcaggg taggaccagg
      721 tctgtgcctg acacaacggt gatgttcttg ggaatagatc tgactttttt aatgccatac
      781 tggttacaca tctttgaagt gtccaagttc gccacatgtt ggaccagaaa ggacccagag
      841 cgaccactgg tggtggccac acagcggctg gtgttgacat tccaggcaca gtaagggtcc
      901 ctggccagga cacagtctac acagaaacgg tactttgtgc agtcggccag agacagctga
      961 accagctgag agcgggagcc agcaaagagc accttcttgc tctgagacag caccagactt
     1021 tccactggct cctggtcaaa cacctgcagt tcctccacca tgtggatcca gggccccagg
     1081 ctcacagcct tcagcagcca gccatctcct gtaccaatga acaacactgt ataggtggca
     1141 ccatcaagcc ctgggaccct gtcggccacc acgtgtgtga agttagtgtt cttcttcaca
     1201 agtagggggc ggcccaaccg aggcttcacc tggtcctcca tcagggggtg cttcttgatg
     1261 aagttgaggg tgttgtccgg cagttccagg gaactggtgt agccattgtc tcggtgccag
     1321 ttgttgatac acgaaccagg ccgagggctg ggtaccgggt cagtatagcg ggcccacttc
     1381 tgggcttgct cactgtactc cttgtaggga ccctcaaaca cttgctggat ctgttccaac
     1441 tggtactcac aaactgcaga caggtccata tcgccccatc gcgcttgaaa aaccccgaag
     1501 aaggtggtgt tgtgccaaga ggcgccccgc agggtgtgca ccgccttcag ctggttgaag
     1561 tagaccttcc agtcaggggc tgagcacacc aaccgagccc tcaggaacgt cgtccatttc
     1621 ttctgcagcg tccgtgctcc ccccatgtca cccttacaga ctctcgccac acgagccacc
     1681 acctgctcgg aatagcagtc atactccact gcccgctcac tgaagaagaa gtagatcttg
     1741 tcatcgtctc ccgtgaagct tcccacactc tcagggacaa aggcagagcc tacaaagtgg
     1801 ggttcattca gccaaaaagc caggtactct gtcttgatgg agtggtgggt ccccatgtat
     1861 cgaaggataa ccggctctgt gcccaggaag ttattgagtg tggctgagta cagctcaccg
     1921 tccacaagga gtccggtgtg acccttagct gggtcatatg ggcatttacc cttcccatcc
     1981 tcaaattctg cacggtccaa ggtgaacgtg agcatgttga tgtaggtgca cttgggctgg
     2041 aaggcatagg tgccgcagac atacagatgg gaggaattgt atggctgaag gaagcggatg
     2101 aagttgaagc attcggtctg gttgctcttc cctttctggg tacattcaat tttcttctca
     2161 gctggagcct cccaagagat cgctccttgc agctccagag cctctacact gaaggcaaac
     2221 agcgcctctc gggcccccac atataaaagg ccagaatgtt ctgtcagggt cagtgtcagg
     2281 aagtcctgga tgcctgtctg ggagaaccgc ctcactactg tgaccagctc cccagaagat
     2341 actgtcttcc ggggcacaag gttccaccac atctcagccc cgatgcccag gccccacagc
     2401 cctgctgcca gcagccagac agcccagtgt ggggccataa gggcgaattc tgcagatatc
     2461 catcacactg gcggccgctc gagcatgcat ctagagggcc caattcgccc tatagtgagt
     2521 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2581 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2641 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     2701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2761 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2881 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2941 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3001 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3061 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3121 taacaaaatt cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg
     3181 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     3241 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     3301 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     3361 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     3421 ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3481 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3541 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3601 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3661 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3721 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatcccac
     3781 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3841 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3901 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3961 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4021 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4081 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4141 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4201 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     4261 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4321 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4381 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4441 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4501 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4561 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4621 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4681 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4741 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4801 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4861 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4921 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4981 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5041 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5101 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5161 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5221 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5881 ggagcctatg gaaaaacgcc agcaacgcgg
//
