LOCUS       sequence-202953-        4383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rosa26 locus targeting vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4383)
  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 4383)
  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
FEATURES             Location/Qualifiers
     source          1..4383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(132..149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(266..811)
                     /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(346..365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1987..2797
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     misc_feature    2311..2960
                     /label=HA-R
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     CDS             complement(3239..4054)
                     /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(3239..4054)
                     /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     3962..3981
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4046..4065
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
       61 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
      121 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
      181 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
      241 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
      301 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
      361 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
      421 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
      481 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
      541 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
      601 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
      661 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
      721 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
      781 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
      841 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
      901 atataagttg tatttcgtcg acggaagagc ggcgcgccaa atctccgttc tgtgagacag
      961 ccgggtacga gtcgtgacgc tggaaggggc aagcgggtgg tgggcaggaa tgcggtccgc
     1021 cctgcagcaa ccggaggggg agggagaagg gagcggaaaa gtctccaccg gacgcggcca
     1081 tggctcgggg gggggggggc agcggaggag cgcttccggc cgacgtctcg tcgctgattg
     1141 gcttcttttc ctcccgccgt gtgtgaaaac acaaatggcg tgttttggtt ggcgtaaggc
     1201 gcctgtcagt taacggcagc cggagtgcgc agccgccggc agcctcgctc tgcccactgg
     1261 gtggggcggg aggtaggtgg ggtgaggcga gctggacgtg cgggcgcggt cggcctctgg
     1321 cggggcgggg grggggaggg agggtcagcg aaagtagctc gcgcgcgagc ggccgcccac
     1381 cctccccttc ctctggggga gtcgttttac ccgccgccgg ccgggcctcg tcgtctgatt
     1441 ggctctcggg gcccagaaaa ctggcccttg ccattggctc gtgttcgtgc aagttgagtc
     1501 catccgccgg ccagcggggg cggcgaggag gcgctcccag gttccggccc tcccctcggc
     1561 cccgcgccgc agagtctggc cgcgcgcccc tgcgcaacgt ggcaggaagc gcgcgctggg
     1621 ggcggggacg ggcagtaggg ctgagcggct gcggggcggg tgcaagcacg tttccgactt
     1681 gagttgcctc aagaggggcg tgctgagcca gacctccatc gcgcactccg gggagtggag
     1741 ggaaggagcg agggctcagt tgggctgttt tggaggcagg aagcacttgc tctcccaaag
     1801 tcgctctgag ttgttatcag taagggagct gcagtggagt aggcggggag aaggccgcac
     1861 ccttctccgg aggggggagg ggagtgttgc aatacctttc tgggagttct ctgctgcctc
     1921 ctggcttctg aggaccgccc tgggcctggg agaatccctt ccccctcttc tttctttaat
     1981 taaagaagat gggcgggagt cttctgggca ggcttaaagg ctaacctggt gtgtgggcgt
     2041 tgtcctgcag gggaattgaa caggtgtaaa attggaggga caagacttcc cacagatttt
     2101 cggttttgtc gggaagtttt ttaatagggg caaataagga aaatgggagg ataggtagtc
     2161 atctggggtt ttatgcagca aaactacagg ttattattgc ttgtgatccg cctcggagta
     2221 ttttccatcg aggtagatta aagacatgct cacccgagtt ttatactctc ctgcttgaga
     2281 tccttactac agtatgaaat tacagtgtcg cgagttagac tatgtaagca gaattttaat
     2341 catttttaaa gagcccagta cttcatatcc atttctcccg ctccttctgc agccttatca
     2401 aaaggtattt tagaacactc attttagccc cattttcatt tattatactg gcttatccaa
     2461 cccctagaca gagcattggc attttccctt tcctgatctt agaagtctga tgactcatga
     2521 aaccagacag attagttaca tacaccacaa atcgaggctg tagctggggc ctcaacactg
     2581 cagttctttt ataactcctt agtacacttt ttgttgatcc tttgccttga tccttaattt
     2641 tcagtgtcta tcacctctcc cgtcaggtgg tgttccacat ttgggcctat tctcagtcca
     2701 gggagtttta caacaataga tgtattgaga atccaaccta aagcttaact ttccactccc
     2761 atgaatgcct ctctcctttt tctccattta taaactgagc tattaaccat taatggtttc
     2821 caggtggatg tctcctcccc caatattacc tgatgtatct tacatattgc caggctgata
     2881 ttttaagaca ttaaaaggta tatttcatta ttgagccaca tggtattgat tactgcttac
     2941 taaaattttg tcattgtaca catctgtaaa aggtggttcc ttttggaatg caaagttcag
     3001 gtgtttgttg tctttcctga cctaaggtct tgtgagcttg tattttttct atttaagcag
     3061 tgctttctct tggactggct tgactcatgg cattctacac gttattgctg gtctaaatgt
     3121 gattttgcca agcttcttca ggacctataa ttttgcttga cttgtagcca aacacaagta
     3181 aaatgattaa gcaacaaatg tatttgtgaa gcttggtcca ccacgctcgg agggcctgtt
     3241 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     3301 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     3361 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     3421 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     3481 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     3541 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     3601 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     3661 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     3721 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     3781 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     3841 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     3901 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     3961 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     4021 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     4081 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     4141 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     4201 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     4261 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     4321 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
     4381 agc
//
