LOCUS       sequence-202939-        7531 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Rosa26 targeting vector for Dre-responsive cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7531)
  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 7531)
  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_RNeoStop_WPRE_bgh_pA
FEATURES             Location/Qualifiers
     source          1..7531
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     8..27
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    57..88
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             89..892
                     /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(152..171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     762..781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          1435..1500
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1630..1650
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2075..2209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2112..2131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2166..2185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2251..2282
                     /label=rox
                     /bound_moiety="Dre recombinase"
     misc_feature    2308..2896
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2361..2381)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2779..2790)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2912..3136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3148..3958
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     misc_feature    3472..4121
                     /label=HA-R
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     CDS             complement(4400..5215)
                     /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(4400..5215)
                     /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     5123..5142
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5207..5226
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5676..5693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5810..6355)
                     /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(5890..5909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgattaac
       61 tttaaataat tggcattatt taaagttaat gggatcggcc attgaacaag atggattgca
      121 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      181 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      241 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      301 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      361 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      421 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      481 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      541 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      601 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca
      661 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
      721 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
      781 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
      841 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct aatagtgaga
      901 tattaagggt tccggatcct cggggacacc aaatatggcg atctcggcct tttcgtttct
      961 tggagctggg acatgtttgc catcgatcca tctaccacca gaacggccgt tagatctgct
     1021 gccaccgttg tttccaccga agaaaccacc gttgccgtaa ccaccacgac ggttgttgct
     1081 aaagaagctg ccaccgccac ggccaccgtt gtagccgccg ttgttgttat tgtagttgct
     1141 actgttattt ctggcacttc ttggttttcc tcttaagtga ggaggaacat aaccattctc
     1201 gttgttgtcg ttgatgctta aattttgcac ttgttcgctc agttcagcca taatatgaaa
     1261 tgcttttctt gttgttctta cggaatacca cttgccacct atcaccacaa ctaacttttt
     1321 cccgttcctc catctctttt atattttttt tctcgaggga tctttgtgaa ggaaccttac
     1381 ttctgtggtg tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat
     1441 ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag
     1501 attccaacct atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac
     1561 ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat
     1621 tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg
     1681 ctaagttttt tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac
     1741 accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc
     1801 tttataagta ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg
     1861 catagagtgt ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt
     1921 tgtaaagggg ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag
     1981 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2041 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2101 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2161 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gacatggtaa
     2221 gtaagcttgg gctgcaggtc gagggaccta taactttaaa taattggcat tatttaaagt
     2281 taatcaggcc tgtttaaaca tttaaataat caacctctgg attacaaaat ttgtgaaaga
     2341 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2401 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2461 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2521 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2581 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2641 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2701 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2761 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2821 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2881 tgggccgcct ccccgcctga aataattccg actgtgcctt ctagttgcca gccatctgtt
     2941 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3001 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3061 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3121 gcggtgggct ctatggttaa ttaaagaaga tgggcgggag tcttctgggc aggcttaaag
     3181 gctaacctgg tgtgtgggcg ttgtcctgca ggggaattga acaggtgtaa aattggaggg
     3241 acaagacttc ccacagattt tcggttttgt cgggaagttt tttaataggg gcaaataagg
     3301 aaaatgggag gataggtagt catctggggt tttatgcagc aaaactacag gttattattg
     3361 cttgtgatcc gcctcggagt attttccatc gaggtagatt aaagacatgc tcacccgagt
     3421 tttatactct cctgcttgag atccttacta cagtatgaaa ttacagtgtc gcgagttaga
     3481 ctatgtaagc agaattttaa tcatttttaa agagcccagt acttcatatc catttctccc
     3541 gctccttctg cagccttatc aaaaggtatt ttagaacact cattttagcc ccattttcat
     3601 ttattatact ggcttatcca acccctagac agagcattgg cattttccct ttcctgatct
     3661 tagaagtctg atgactcatg aaaccagaca gattagttac atacaccaca aatcgaggct
     3721 gtagctgggg cctcaacact gcagttcttt tataactcct tagtacactt tttgttgatc
     3781 ctttgccttg atccttaatt ttcagtgtct atcacctctc ccgtcaggtg gtgttccaca
     3841 tttgggccta ttctcagtcc agggagtttt acaacaatag atgtattgag aatccaacct
     3901 aaagcttaac tttccactcc catgaatgcc tctctccttt ttctccattt ataaactgag
     3961 ctattaacca ttaatggttt ccaggtggat gtctcctccc ccaatattac ctgatgtatc
     4021 ttacatattg ccaggctgat attttaagac attaaaaggt atatttcatt attgagccac
     4081 atggtattga ttactgctta ctaaaatttt gtcattgtac acatctgtaa aaggtggttc
     4141 cttttggaat gcaaagttca ggtgtttgtt gtctttcctg acctaaggtc ttgtgagctt
     4201 gtattttttc tatttaagca gtgctttctc ttggactggc ttgactcatg gcattctaca
     4261 cgttattgct ggtctaaatg tgattttgcc aagcttcttc aggacctata attttgcttg
     4321 acttgtagcc aaacacaagt aaaatgatta agcaacaaat gtatttgtga agcttggtcc
     4381 accacgctcg gagggcctgt tagaaaaact catcgagcat caaatgaaac tgcaatttat
     4441 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     4501 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     4561 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     4621 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
     4681 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     4741 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     4801 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     4861 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     4921 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     4981 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     5041 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     5101 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     5161 tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg ctcataacac
     5221 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     5281 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     5341 cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg
     5401 caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg
     5461 gctccctcac tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac
     5521 cttcttcacg aggcagacct cagcgctagc ggagtgtata ctggcttact atgttggcac
     5581 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     5641 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     5701 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     5761 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     5821 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     5881 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     5941 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     6001 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     6061 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     6121 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     6181 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     6241 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     6301 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     6361 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     6421 agcacctgaa gtcagcccca tacgatataa gttgtatttc gtcgacggaa gagcggcgcg
     6481 ccaaatctcc gttctgtgag acagccgggt acgagtcgtg acgctggaag gggcaagcgg
     6541 gtggtgggca ggaatgcggt ccgccctgca gcaaccggag ggggagggag aagggagcgg
     6601 aaaagtctcc accggacgcg gccatggctc gggggggggg gggcagcgga ggagcgcttc
     6661 cggccgacgt ctcgtcgctg attggcttct tttcctcccg ccgtgtgtga aaacacaaat
     6721 ggcgtgtttt ggttggcgta aggcgcctgt cagttaacgg cagccggagt gcgcagccgc
     6781 cggcagcctc gctctgccca ctgggtgggg cgggaggtag gtggggtgag gcgagctgga
     6841 cgtgcgggcg cggtcggcct ctggcggggc gggggagggg agggagggtc agcgaaagta
     6901 gctcgcgcgc gagcggccgc ccaccctccc cttcctctgg gggagtcgtt ttacccgccg
     6961 ccggccgggc ctcgtcgtct gattggctct cggggcccag aaaactggcc cttgccattg
     7021 gctcgtgttc gtgcaagttg agtccatccg ccggccagcg ggggcggcga ggaggcgctc
     7081 ccaggttccg gccctcccct cggccccgcg ccgcagagtc tggccgcgcg cccctgcgca
     7141 acgtggcagg aagcgcgcgc tgggggcggg gacgggcagt agggctgagc ggctgcgggg
     7201 cgggtgcaag cacgtttccg acttgagttg cctcaagagg ggcgtgctga gccagacctc
     7261 catcgcgcac tccggggagt ggagggaagg agcgagggct cagttgggct gttttggagg
     7321 caggaagcac ttgctctccc aaagtcgctc tgagttgtta tcagtaaggg agctgcagtg
     7381 gagtaggcgg ggagaaggcc gcacccttct ccggaggggg gaggggagtg ttgcaatacc
     7441 tttctgggag ttctctgctg cctcctggct tctgaggacc gccctgggcc tgggagaatc
     7501 ccttccccct cttctttctt taattaagct c
//
