LOCUS       sequence-118631-        6060 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 6060)
  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 6060)
  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-Sema4a-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2157..2176)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2158..2176)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2183..2200)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2183..2199)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2192..2214)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2340..2768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2427..2446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2637..2658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3112..3906
                     /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(3166..3185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3776..3795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3924..3992,3993..4784)
                     /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(4142..4161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4955..5543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5444..5463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5697..5714
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5831..5852
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5867..5872,5873..5890,5891..5897)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5905..5921
                     /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     5910..5932
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5929..5945
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5929..5945
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5963..5981
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5963..5980
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 agagtgagca ctcccaggag cacgatggcc aggaggacgg taacgatgag aaaatggggc
       61 cagtaggacc gctgggcagc catggaagct ccgcctccga ccctggtcag cgggacctgc
      121 acacgctcac ggggaacgcc cgccagctca gggtccagcg ccaggggctg gtcctggctg
      181 tctacccaat aggagaccac agggtatgag tagccgttct cagtcgccac acactggtag
      241 aggcccccga caccatcctg cggcagcagc aagagggagc cattgtagac ggtagcagag
      301 gcttctgaga ttttggctcg gccatgactc cagtggtaag aggccagtgc tgacaggtgg
      361 gggcagggca gctccaggat ggagttgggg actgtcagga cttctttaat tagttgaggg
      421 gggctctgac gccgggggct cctggccatg gggccacggg tgcataccca ctccgggttg
      481 ccgcgttcca tgtcctgctt ccaaggcttg gtagagccag acagaaggct gcagagtctt
      541 gattcagggt cccaggcaca gtgagggtcc ctggcaagca cacagtccac acagctctcg
      601 tagacactgc aattggccct gggaactctc cagatgcctc cagagaagcc tgcaaacact
      661 gcaccctggg cgggggccag ctgcaggttt cgaacaggct cagagtcagg gctcagctga
      721 atctcctcca cgagataagc actgctgtcc tgaggcacca cagccttgtg cagggacccc
      781 gtggaggtac ccagatacat gaccacatgg ctgctcccat caaggccccg agctgactcc
      841 acagcaagcc gtgtgtactc cacaccagac ttcaccagca ggggtgttcc taccacgtgc
      901 tcatccatca gaaaatggtc cttcatgaag gtcaaggctt tgtcagagga ggggcccatg
      961 gagcaactgc ctggcctcgg gctgacctct gagccccggt aagtggtcca gcgggaggtc
     1021 tccttgttca gctccttgta cttcccttta aagactcgct caatgtccgt gagagagaag
     1081 gcacagactg ctgagctcct ggtcccgcca acctgccact gggaggtaaa gactgcgtag
     1141 atgcgggaaa cagagggaga atcggcgggc agcaggaccg cgtggcggat gatgttgaat
     1201 ggcagctgcc ctggctgagc gcagagcaac tgggctttga ggaaggtggt ccacttcttc
     1261 tgcagcagct tttcaccgcc cacgtcgttc ttgcagactt gagccaccct ggatatatac
     1321 agctcttcaa agaagtcaaa ctcgctggct gtctcctcaa agaagaaata gacgacctgg
     1381 gtggatggaa tggctgccac gaaggaggca tccgcgtgca gccagcgtaa gaagatgtca
     1441 gtcttgagaa caggctggga tcccagtgtc cgcatcagga tgggctcgct gcccaggaag
     1501 ttgttcatgg tgccggaata aagcatccca tcgaccaaga cagctgtgtg cttgtgaaca
     1561 gggtcaaagg ggctttggcc cttcccgtcc atgaccttgt ctatcaagat gggcaacagg
     1621 agggaatctt ggagttcaat gaaggtacag gcagggctga aggcaaaggt cccacaggca
     1681 tagaggtgag tagcattgta agagaccagg actcgaatga agttgaaaca ctgtgtctca
     1741 ttgctcttct tcttaaaggc acattcggtc ttttttctct cactggctgg ccagggtatc
     1801 atgttcttta gccttgggat tcctgggttc tggatattca aggccaggac ggcctctcga
     1861 gcccccacat agagagtgtt gccatcgtca ctcaggagca gcgtgtcaaa gtctcggagg
     1921 cctttttgtt ggaagaagct gagggccctg tgcccgtctc cagcatggta tttgactctg
     1981 ggcatgggcc cctgaccacc agtcccagaa gcaggtggca gtgatggcag caggaagagt
     2041 tggaagaaaa aaacacgcag gagactccat gagtcctggc ccagggaaag ggcgaattct
     2101 gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc aattcgccct
     2161 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2221 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2281 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2341 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2401 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2461 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2521 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2581 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2641 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2701 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2761 cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     2821 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     2881 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag
     2941 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     3001 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
     3061 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3121 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3181 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3241 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3301 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3361 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3421 tcatcccacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3481 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3541 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3601 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3661 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3721 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3781 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3841 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3901 ttctgaattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3961 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4021 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4081 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4141 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4201 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4261 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4321 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4441 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5641 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5701 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5761 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5821 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5881 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5941 atgaccatga ttacgccaag ctatttaggt gacactatag aatactcaag ctatgcatca
     6001 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attcgccctt
//
