LOCUS       sequence-202870-        7609 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Rosa26 targeting vector for FLP-responsive cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7609)
  AUTHORS   Lusk SJ, McKinney A, Hunt PJ, Fahey PG, Patel J, Chang A, Sun JJ, 
            Martinez VK, Zhu PJ, Egbert JR, Allen G, Jiang X, Arenkiel BR, 
            Tolias AS, Costa-Mattioli M, Ray RS
  TITLE     A CRISPR toolbox for generating intersectional genetic mouse models 
            for functional, molecular, and anatomical circuit mapping.
  JOURNAL   BMC Biol. 2022 Jan 28;20(1):28. doi: 10.1186/s12915-022-01227-0.
  PUBMED    35086530
REFERENCE   2  (bases 1 to 7609)
  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"; journalName: "BMC Biol.";
            date: "2022-01-28"; volume: "20(1):28. doi"; pages: " 
            10.1186/s12915-022-01227-0"
COMMENT     Sequence Label: 
            p15a_Kan_MLS_Rosa26_Short_CRISPRDel_CAG_FNeoStop_WPRE_bgh_pA
FEATURES             Location/Qualifiers
     source          1..7609
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     80..99
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    129..162
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             166..969
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(229..248)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     839..858
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          1512..1577
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1707..1727
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2152..2286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2189..2208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2243..2262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2328..2361
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    2387..2975
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2440..2460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2858..2869)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2991..3215
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3227..4037
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     misc_feature    3551..4200
                     /label=HA-R
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     CDS             complement(4479..5294)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4479..5294)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5202..5221
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5286..5305
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5755..5772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5889..6434)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5969..5988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
       61 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
      121 gaattgatga agttcctatt ctctagaaag tataggaact tcaccatggg atcggccatt
      181 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
      241 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
      301 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
      361 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
      421 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
      481 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
      541 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
      601 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
      661 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat
      721 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
      781 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
      841 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
      901 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
      961 ttcttctaat agtgagatat taagggttcc ggatcctcgg ggacaccaaa tatggcgatc
     1021 tcggcctttt cgtttcttgg agctgggaca tgtttgccat cgatccatct accaccagaa
     1081 cggccgttag atctgctgcc accgttgttt ccaccgaaga aaccaccgtt gccgtaacca
     1141 ccacgacggt tgttgctaaa gaagctgcca ccgccacggc caccgttgta gccgccgttg
     1201 ttgttattgt agttgctact gttatttctg gcacttcttg gttttcctct taagtgagga
     1261 ggaacataac cattctcgtt gttgtcgttg atgcttaaat tttgcacttg ttcgctcagt
     1321 tcagccataa tatgaaatgc ttttcttgtt gttcttacgg aataccactt gccacctatc
     1381 accacaacta actttttccc gttcctccat ctcttttata ttttttttct cgagggatct
     1441 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
     1501 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
     1561 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
     1621 cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta
     1681 ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg
     1741 actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg
     1801 cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg
     1861 aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt
     1921 ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta
     1981 cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga
     2041 ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2101 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2161 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2221 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2281 tggatctgac atggtaagta agcttgggct gcaggtcgag ggacctagaa gttcctattc
     2341 tctagaaagt ataggaactt catcaggcct gtttaaacat ttaaataatc aacctctgga
     2401 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2461 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2521 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2581 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2641 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2701 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2761 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2821 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2881 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2941 gacgagtcgg atctcccttt gggccgcctc cccgcctgaa ataattccga ctgtgccttc
     3001 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3061 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3121 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3181 tagcaggcat gctggggatg cggtgggctc tatggttaat taaagaagat gggcgggagt
     3241 cttctgggca ggcttaaagg ctaacctggt gtgtgggcgt tgtcctgcag gggaattgaa
     3301 caggtgtaaa attggaggga caagacttcc cacagatttt cggttttgtc gggaagtttt
     3361 ttaatagggg caaataagga aaatgggagg ataggtagtc atctggggtt ttatgcagca
     3421 aaactacagg ttattattgc ttgtgatccg cctcggagta ttttccatcg aggtagatta
     3481 aagacatgct cacccgagtt ttatactctc ctgcttgaga tccttactac agtatgaaat
     3541 tacagtgtcg cgagttagac tatgtaagca gaattttaat catttttaaa gagcccagta
     3601 cttcatatcc atttctcccg ctccttctgc agccttatca aaaggtattt tagaacactc
     3661 attttagccc cattttcatt tattatactg gcttatccaa cccctagaca gagcattggc
     3721 attttccctt tcctgatctt agaagtctga tgactcatga aaccagacag attagttaca
     3781 tacaccacaa atcgaggctg tagctggggc ctcaacactg cagttctttt ataactcctt
     3841 agtacacttt ttgttgatcc tttgccttga tccttaattt tcagtgtcta tcacctctcc
     3901 cgtcaggtgg tgttccacat ttgggcctat tctcagtcca gggagtttta caacaataga
     3961 tgtattgaga atccaaccta aagcttaact ttccactccc atgaatgcct ctctcctttt
     4021 tctccattta taaactgagc tattaaccat taatggtttc caggtggatg tctcctcccc
     4081 caatattacc tgatgtatct tacatattgc caggctgata ttttaagaca ttaaaaggta
     4141 tatttcatta ttgagccaca tggtattgat tactgcttac taaaattttg tcattgtaca
     4201 catctgtaaa aggtggttcc ttttggaatg caaagttcag gtgtttgttg tctttcctga
     4261 cctaaggtct tgtgagcttg tattttttct atttaagcag tgctttctct tggactggct
     4321 tgactcatgg cattctacac gttattgctg gtctaaatgt gattttgcca agcttcttca
     4381 ggacctataa ttttgcttga cttgtagcca aacacaagta aaatgattaa gcaacaaatg
     4441 tatttgtgaa gcttggtcca ccacgctcgg agggcctgtt agaaaaactc atcgagcatc
     4501 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     4561 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     4621 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     4681 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     4741 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     4801 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     4861 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     4921 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     4981 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     5041 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     5101 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     5161 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     5221 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt
     5281 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     5341 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     5401 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac
     5461 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac
     5521 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc
     5581 tggtatgagt cagcaacacc ttcttcacga ggcagacctc agcgctagcg gagtgtatac
     5641 tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa
     5701 aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct
     5761 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg
     5821 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
     5881 aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat
     5941 cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc
     6001 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
     6061 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
     6121 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
     6181 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
     6241 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
     6301 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
     6361 tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac
     6421 caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc
     6481 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtatttcg
     6541 tcgacggaag agcggcgcgc caaatctccg ttctgtgaga cagccgggta cgagtcgtga
     6601 cgctggaagg ggcaagcggg tggtgggcag gaatgcggtc cgccctgcag caaccggagg
     6661 gggagggaga agggagcgga aaagtctcca ccggacgcgg ccatggctcg gggggggggg
     6721 ggcagcggag gagcgcttcc ggccgacgtc tcgtcgctga ttggcttctt ttcctcccgc
     6781 cgtgtgtgaa aacacaaatg gcgtgttttg gttggcgtaa ggcgcctgtc agttaacggc
     6841 agccggagtg cgcagccgcc ggcagcctcg ctctgcccac tgggtggggc gggaggtagg
     6901 tggggtgagg cgagctggac gtgcgggcgc ggtcggcctc tggcggggcg ggggrgggga
     6961 gggagggtca gcgaaagtag ctcgcgcgcg agcggccgcc caccctcccc ttcctctggg
     7021 ggagtcgttt tacccgccgc cggccgggcc tcgtcgtctg attggctctc ggggcccaga
     7081 aaactggccc ttgccattgg ctcgtgttcg tgcaagttga gtccatccgc cggccagcgg
     7141 gggcggcgag gaggcgctcc caggttccgg ccctcccctc ggccccgcgc cgcagagtct
     7201 ggccgcgcgc ccctgcgcaa cgtggcagga agcgcgcgct gggggcgggg acgggcagta
     7261 gggctgagcg gctgcggggc gggtgcaagc acgtttccga cttgagttgc ctcaagaggg
     7321 gcgtgctgag ccagacctcc atcgcgcact ccggggagtg gagggaagga gcgagggctc
     7381 agttgggctg ttttggaggc aggaagcact tgctctccca aagtcgctct gagttgttat
     7441 cagtaaggga gctgcagtgg agtaggcggg gagaaggccg cacccttctc cggagggggg
     7501 aggggagtgt tgcaatacct ttctgggagt tctctgctgc ctcctggctt ctgaggaccg
     7561 ccctgggcct gggagaatcc cttccccctc ttctttcttt aattaagct
//
