LOCUS       sequence-118658-        6107 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 6107)
  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 6107)
  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-Sema4d-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..6107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2204..2223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2205..2223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2230..2247)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2230..2246)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2239..2261)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2387..2815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2474..2493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2684..2705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3159..3953
                     /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(3213..3232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3823..3842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3971..4039,4040..4831)
                     /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(4189..4208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5002..5590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5491..5510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5744..5761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5878..5899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5914..5919,5920..5937,5938..5944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5952..5968
                     /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     5957..5979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5976..5992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5976..5992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6010..6028
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6010..6027
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ccatctttgg ttcacaggat gtgccggagg aggacttggg aggtagggtg gcggctctgg
       61 gggaggttgc ccacagagcc ggggtagggg gagaggaccc ctgggtagat gcaaccggca
      121 ttttggatgt gatcttacta ccttctgtct gagcatcctc tgaggtaggt gagggggggg
      181 tccggggtac catcttcact tccagaacgt gcttggccag cagctgggag accgttttat
      241 tcctcaccct ttcctctgac aggcactggt acacgccgct gtctccgtcc gacaggttga
      301 agatgagcag gtgcttcctg cccacaaagc cgtacttggg acttgcggcc ttcaactcgc
      361 cattctggaa cttccatacc acccgggcta ggttggactt ttggaaacat ttgagttccg
      421 ctgtgccgcc gtgcttgaaa aaatgctggt tgaaactttc tttactctta tccaggcatg
      481 aggatgtgtc accgctcatg tcctgaatcc agcccctgct ggaggcctct tcctggtgca
      541 gggtaacaca ggccttgatg gctgggctcc aggcacagta ggggtcccgt gctaacacac
      601 agtcttcaca gctaccgtgc ttttcgcaga atgccagggg cgcttggacc actccagagt
      661 tggagcctgc atagacaaac ttcctcccct tctttgacga tagcagcaga gttaggaccg
      721 gttcagagtc ccggaagagt tgggtctcct cgatgacatg cacctctttt gtgaggatga
      781 ctgctttatg cagagctccc cggtctgtgc tgatgaacat gacgtcgtag aaagtcccat
      841 ccagggcctg ggtcctgtct accactatct gggtgtagtt tacatctttt ttgatcagct
      901 tgggtctgtt gtctatcggg gtcactgagt catccatcaa agggtggtct tttacaaact
      961 gcagtgtttt gtctgggaga ttcaaggagc tggtgtagtt ggctgcccgg gcctcactgt
     1021 cgatacacgc tccaggtcgg ggagtgggca ctgggccatt gtagcgcacc cacttggtgt
     1081 gagactgctc cactgtggca ctctgcatgt actttccacg ggagaagact gcctccaccg
     1141 tggccagtgt gtaggcgcac accgctgaca gacccacatt gttcagctgt ggggtgaaga
     1201 ccgcatagaa cacaggctcc ttgaggcccg gggccctcag cacaaacaca tcctgaagta
     1261 tgttgaagac caggccactg tctggcttgg agcagatcag cctggccttt aggaaggagg
     1321 tccacttttt ttgcaaagtc cgcaggccgc cctggtcgcc cttgcacacc ctggcaactc
     1381 gcgggatcat caacttgaag acgaattcgt actccacgga tacctccgta aaaaagaagt
     1441 agaccttgtc atcttcaccc tccggaccat ctgggctttt ctggatcacg tcagcaaaga
     1501 cgaagctagg ctcgttcagc cacgggatgg catactccgt cctcaaggga ctgtgggaag
     1561 agtttcgaga gatgatgggt tcactgccca agaaattata ggacgttcca gagtagagct
     1621 cgcccccaac catgactgat gtgtagctgt gggcggggtc gaaggggcat cttcctttgc
     1681 caacttcact tttccccaga aacttgaagg atgtcaagtt caggtggtca caggtgggct
     1741 ggaacgcatt ggtcccacac acatagaggg aagtgctgct tagtggctgt agtactcgaa
     1801 tgtagtttag gcattccgtc tgctttgatt tccccttctc tgcacacttg gattttttgt
     1861 cttcagagac cttccaatat acctcatgtt gcttctcaga gatgttcagc gcattcactg
     1921 caaagactgc ttcccgggcg cctacataca gagtgtcttt gtcctcactc atcagcaagg
     1981 ccgagtagtt aaagatgcct ggcttgtgaa actgcaccag acctacctct ccatgttccc
     2041 aggtgagccg aggcacaggt gcaaatgcca ccgcggttct caacactacc accagggcca
     2101 agaacagccc cctaacgggg gcacacatcc tcataagggc gaattctgca gatatccatc
     2161 acactggcgg ccgctcgagc atgcatctag agggcccaat tcgccctata gtgagtcgta
     2221 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2281 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2341 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     2401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2821 aaaattcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga
     2881 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     2941 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     3001 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     3061 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     3121 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3181 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3241 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3301 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3361 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3421 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tcccaccttg
     3481 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3541 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3601 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3661 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3721 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3781 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3841 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3901 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattgaaa
     3961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4201 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4261 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4321 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4381 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4441 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4501 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4681 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4801 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4861 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4921 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4981 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5041 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5101 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5161 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5221 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5281 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5341 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5401 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5461 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5521 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5581 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5641 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5701 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5761 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5821 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5881 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     5941 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6001 cgccaagcta tttaggtgac actatagaat actcaagcta tgcatcaagc ttggtaccga
     6061 gctcggatcc actagtaacg gccgccagtg tgctggaatt cgccctt
//
