LOCUS       sequence-118656-        5995 bp    DNA     circular SYN 15-APR-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 5995)
  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 5995)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Sema4b-ISH-2kb
FEATURES             Location/Qualifiers
     source          1..5995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             850..861
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2092..2111)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2093..2111)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2118..2135)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2118..2134)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2127..2149)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2275..2703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2362..2381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2572..2593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3047..3841
                     /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(3101..3120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3711..3730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3859..3927,3928..4719)
                     /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(4077..4096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4890..5478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5379..5398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5632..5649
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5766..5787
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5802..5807,5808..5825,5826..5832)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5840..5856
                     /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     5845..5867
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5864..5880
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5864..5880
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5898..5916
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5898..5915
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tgcactcact cgtgatgtgt tgataatgac tggggtacca tcacgctggt tcttttggtc
       61 cattaccccc tcctccatca cctctgggca gtagctggcc acaagctgct ggaatccttc
      121 ttctatcgac caacactgga acacccccaa accctgctgg ctgcccacca gcagcaggtc
      181 cccggtgggt aacacgcggc aggaggcaga ggcattgact ggggctccat tgtgcaccca
      241 gagccgagtg gccaggtttg agaggagtgg gcaggccagg gtgttcactg tgtttggttg
      301 gatctggact tgtttacatg gcttacctgg cacaagaaac cgggccttgt atgaggaatt
      361 cttgcagagt tccttgacac tggcaccctc aatgtcctgg gtccatggcc tggaggccag
      421 atcaggctgg tagaggctag cgagcctgca ggcagagcca gtccaggcgc agtaggggtc
      481 tcgagccagg aggcagtctc cacaggttgg gtacaggctg cagttggcta cgggcacttg
      541 caccaccccg gaatgggagg aggcatacaa cagtccccca tggctgtcca agagcaggtt
      601 ctgcacaggc tgtccttgag ggaagatctg cagctcctca atgatgtgga ctctggagct
      661 cagggtcact gctttgtgca ggcggccatc accagtgccc agaaatagga catcataagt
      721 gctgtgcagg ccaggcacac ggtgcacagc cacacgctgg tagcgggctc tgggctgcag
      781 cagcagcagg cgactgcgga cctgcccatc catcaagaag tgatccttga ggaagttcag
      841 cactcggtct gggagctgca gggacgagtt gatcttccgt tcccgggcac tgttggtaat
      901 gcacgctccc ggccgcggtg tgggcacctg gtgggtctcg gtataccact gctgtgtctc
      961 tctgtttact ttcttgtaca ggccgtcaaa ggccttctgc acatcattca tggtgaagac
     1021 gcagatggca gagccttctg tggtccctct gtgccactgg gaggtaaaga ccccatagaa
     1081 aagggtcttg cgccaatcct gagggttggg gttcagggtg aagacatctt gtagcacgtt
     1141 aaagggaaag ccatcatcag gccgggagca caggagctga gccttgagaa aggaggccca
     1201 gcgttgctgc aacacccgct ctccaccctc atcgccctta cagactcggg caactcggga
     1261 cacgatggtg ttctcaaaga actcaaactc ctggcccgtc tcgctgaaga agaagtagat
     1321 cttatcatca tcacctatgg ggctgcccag gctctcgggg acgtaggccg aggccacaaa
     1381 ggcagggtct tgtagccagt tgagggagct ctcagtcttg gtggggcggg aactctggct
     1441 ccgggaaatg gctgggtcgt ttccctggaa gctactgact gttccagtgt acagctcacc
     1501 atcaaccacc agagccgtgg acttgaagtt ggggtcaaag ggacaacgac ccttgccatc
     1561 ctccagaatg acattgccgg cctcatcttg ggctaaagta aagctcgcta tgtgaatgta
     1621 agcacacagg gggctgaagg cggccgtgcc acaggtgagc aggtggctgc tgttgagtgg
     1681 caggaggatc ttgatgtagt tttgacagtc acgctttggg tccttgccct tgaagctgca
     1741 ctgctgcttc ctgtcagcat ctgcactcca cagcagctct tggtactccc cgcctggcaa
     1801 gaagctgagg ttgctgttaa gtgcaaagag ggcctctcgg gcccccacat acagcgtctt
     1861 gccatcctgg ctcagcagaa gggccgtgta gttggagatg ttttcagctt caaattttct
     1921 aatcagcctt tcttcagagc ccagcggcac actgatccgg gggccgagcg cccgcgtagt
     1981 cgtcgtccgc agcagcagca gtagcagcag cagcagcagc gcaagggcga attctgcaga
     2041 tatccatcac actggcggcc gctcgagcat gcatctagag ggcccaattc gccctatagt
     2101 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2161 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2221 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     2281 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2341 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2401 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2461 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2521 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2581 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2641 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2701 attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag
     2761 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     2821 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     2881 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     2941 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     3001 caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3061 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3121 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3181 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3241 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3301 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3361 ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3421 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3481 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3541 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3601 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3661 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3721 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3781 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3841 aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3961 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4021 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4081 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4141 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4261 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4381 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4441 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4561 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4681 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5521 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5581 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5641 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5701 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5761 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     5821 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     5881 catgattacg ccaagctatt taggtgacac tatagaatac tcaagctatg catcaagctt
     5941 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccctt
//
