LOCUS       sequence-199264-        8323 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  IfgMosaic acceptor vector for Classical or Cas9 Gene targeting of 
            the Rosa26 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Pontes-Quero S, Heredia L, Casquero-Garcia V, Fernandez-Chacon M, 
            Luo W, Hermoso A, Bansal M, Garcia-Gonzalez I, Sanchez-Munoz MS, 
            Perea JR, Galiana-Simal A, Rodriguez-Arabaolaza I, Del Olmo-Cabrera 
            S, Rocha SF, Criado-Rodriguez LM, Giovinazzo G, Benedito R
  TITLE     Dual ifgMosaic: A Versatile Method for Multispectral and 
            Combinatorial Mosaic Gene-Function Analysis.
  JOURNAL   Cell. 2017 Aug 10;170(4):800-814.e18. doi: 
            10.1016/j.cell.2017.07.031.
  PUBMED    28802047
REFERENCE   2  (bases 1 to 8323)
  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.cell.2017.07"; journalName: "Cell"; date: "2017-08-10- 
            10"; volume: "170"; issue: "4"; pages: "800-814.e18"
COMMENT     Sequence Label: pRosa26-4xInsulator (JP6)
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    32..4372
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(4423..4439)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4423..4439)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4471..4490)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4472..4490)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4500..4517)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4500..4516)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4509..4531)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4658..5086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4745..4764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4955..4976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5137..5241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5242..5310,5311..6102)
                     /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(5460..5479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6273..6861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6762..6781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7015..7032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7149..7170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7185..7190,7191..7208,7209..7215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7223..7239
                     /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     7228..7250
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7247..7263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7247..7263
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7282..7302
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7284..7302
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     7332..7348
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cttaattaac ggcggcggag tatcgaattc gctagaagat gggcgggagt cttctgggca
       61 ggcttaaagg ctaacctggt gtgtgggcgt tgtcctgcag gggaattgaa caggtgtaaa
      121 attggaggga caagacttcc cacagatttt cggttttgtc gggaagtttt ttaatagggg
      181 caaataagga aaatgggagg ataggtagtc atctggggtt ttatgcagca aaactacagg
      241 ttattattgc ttgtgatccg cctcggagta ttttccatcg aggtagatta aagacatgct
      301 cacccgagtt ttatactctc ctgcttgaga tccttactac agtatgaaat tacagtgtcg
      361 cgagttagac tatgtaagca gaattttaat catttttaaa gagcccagta cttcatatcc
      421 atttctcccg ctccttctgc agccttatca aaaggtattt tagaacactc attttagccc
      481 cattttcatt tattatactg gcttatccaa cccctagaca gagcattggc attttccctt
      541 tcctgatctt agaagtctga tgactcatga aaccagacag attagttaca tacaccacaa
      601 atcgaggctg tagctggggc ctcaacactg cagttctttt ataactcctt agtacacttt
      661 ttgttgatcc tttgccttga tccttaattt tcagtgtcta tcacctctcc cgtcaggtgg
      721 tgttccacat ttgggcctat tctcagtcca gggagtttta caacaataga tgtattgaga
      781 atccaaccta aagcttaact ttccactccc atgaatgcct ctctcctttt tctccattta
      841 taaactgagc tattaaccat taatggtttc caggtggatg tctcctcccc caatattacc
      901 tgatgtatct tacatattgc caggctgata ttttaagaca ttaaaaggta tatttcatta
      961 ttgagccaca tggtattgat tactgcttac taaaattttg tcattgtaca catctgtaaa
     1021 aggtggttcc ttttggaatg caaagttcag gtgtttgttg tctttcctga cctaaggtct
     1081 tgtgagcttg tattttttct atttaagcag tgctttctct tggactggct tgactcatgg
     1141 cattctacac gttattgctg gtctaaatgt gattttgcca agcttcttca ggacctataa
     1201 ttttgcttga cttgtagcca aacacaagta aaatgattaa gcaacaaatg tatttgtgaa
     1261 gcttggtttt taggttgttg tgttgtgtgt gcttgtgctc tataataata ctatccaggg
     1321 gctggagagg tggctcggag ttcaagagca cagactgctc ttccagaagt cctgagttca
     1381 attcccagca accacatggt ggctcacaac catctgtaat gggatctgat gccctcttct
     1441 ggtgtgtctg aagaccacaa gtgtattcac attaaataaa taaatcctcc ttcttcttct
     1501 tttttttttt tttaaagaga atactgtctc cagtagaatt tactgaagta atgaaatact
     1561 ttgtgtttgt tccaatatgg tagccaataa tcaaattact ctttaagcac tggaaatgtt
     1621 accaaggaac taatttttat ttgaagtgta actgtggaca gaggagccat aactgcagac
     1681 ttgtgggata cagaagacca atgcagactt taatgtcttt tctcttacac taagcaataa
     1741 agaaataaaa attgaacttc tagtatccta tttgtttaaa ctgctagctt tacttaactt
     1801 ttgtgcttca tctatacaaa gctgaaagct aagtctgcag ccattactaa acatgaaagc
     1861 aagtaatgat aattttggat ttcaaaaatg tagggccaga gtttagccag ccagtggtgg
     1921 tgcttgcctt tatgccttta atcccagcac tctggaggca gagacaggca gatctctgag
     1981 tttgagccca gcctggtcta cacatcaagt tctatctagg atagccagga atacacacag
     2041 aaaccctgtt ggggaggggg gctctgagat ttcataaaat tataattgaa gcattcccta
     2101 atgagccact atggatgtgg ctaaatccgt ctacctttct gatgagattt gggtattatt
     2161 ttttctgtct ctgctgttgg ttgggtcttt tgacactgtg ggctttcttt aaagcctcct
     2221 tcctgccatg tggtctcttg tttgctacta acttcccatg gcttaaatgg catggctttt
     2281 tgccttctaa gggcagctgc tgagatttgc agcctgattt ccagggtggg gttgggaaat
     2341 ctttcaaaca ctaaaattgt cctttaattt tttttttaaa aaatgggtta tataataaac
     2401 ctcataaaat agttatgagg agtgaggtgg actaatatta aatgagtccc tcccctataa
     2461 aagagctatt aaggcttttt gtcttatact taactttttt tttaaatgtg gtatctttag
     2521 aaccaagggt cttagagttt tagtatacag aaactgttgc atcgcttaat cagattttct
     2581 agtttcaaat ccagagaatc caaattcttc acagccaaag tcaaattaag aatttctgac
     2641 ttttaatgtt aatttgctta ctgtgaatat aaaaatgata gcttttcctg aggcagggtc
     2701 tcactatgta tctctgcctg atctgcaaca agatatgtag actaaagttc tgcctgcttt
     2761 tgtctcctga atactaaggt taaaatgtag taatactttt ggaacttgca ggtcagattc
     2821 ttttataggg gacacactaa gggagcttgg gtgatagttg gtaaaatgtg tttcaagtga
     2881 tgaaaacttg aattattatc accgcaacct actttttaaa aaaaaaagcc aggcctgtta
     2941 gagcatgctt aagggatccc taggacttgc tgagcacaca agagtagtta cttggcaggc
     3001 tcctggtgag agcatatttc aaaaaacaag gcagacaacc aagaaactac agttaaggtt
     3061 acctgtcttt aaaccatctg catatacaca gggatattaa aatattccaa ataatatttc
     3121 attcaagttt tcccccatca aattgggaca tggatttctc cggtgaatag gcagagttgg
     3181 aaactaaaca aatgttggtt ttgtgatttg tgaaattgtt ttcaagtgat agttaaagcc
     3241 catgagatac agaacaaagc tgctatttcg aggtctcttg gtttatactc agaagcactt
     3301 ctttgggttt ccctgcacta tcctgatcat gtgctaggcc taccttaggc tgattgttgt
     3361 tcaaataaac ttaagtttcc tgtcaggtga tgtcatatga tttcatatat caaggcaaaa
     3421 catgttatat atgttaaaca tttgtactta atgtgaaagt taggtctttg tgggtttgat
     3481 ttttaatttt caaaacctga gctaaataag tcatttttac atgtcttaca tttggtggaa
     3541 ttgtataatt gtggtttgca ggcaagactc tctgacctag taaccctacc tatagagcac
     3601 tttgctgggt cacaagtcta ggagtcaagc atttcacctt gaagttgaga cgttttgtta
     3661 gtgtatacta gtttatatgt tggaggacat gtttatccag aagatattca ggactatttt
     3721 tgactgggct aaggaattga ttctgattag cactgttagt gagcattgag tggcctttag
     3781 gcttgaattg gagtcacttg tatatctcaa ataatgctgg ccttttttaa aaagcccttg
     3841 ttctttatca ccctgttttc tacataattt ttgttcaaag aaatacttgt ttggatctcc
     3901 ttttgacaac aatagcatgt tttcaagcca tatttttttt cctttttttt tttttttttg
     3961 gtttttcgag acagggtttc tctgtatagc cctggctgtc ctggaactca ctttgtagac
     4021 caggctggcc tcgaactcag aaatccgcct gcctctgcct cctgagtgcc gggattaaag
     4081 gcgtgcacca ccacgcctgg ctaagttgga tattttgtta tataactata accaatacta
     4141 actccactgg gtggattttt aattcagtca gtagtcttaa gtggtcttta ttggcccttc
     4201 attaaaatct actgttcact ctaacagagg ctgttggtac tagtggcact taagcaactt
     4261 cctacggata tactagcaga ttaagggtca gggatagaaa ctagtctagc gttttgtata
     4321 cctaccagct ttatactacc ttgttctgat agaaatattt caggacatct agcttatcga
     4381 taccgtcgac gttcgaacac gtggaattcc tgcagcccgg gggatccact agttctagag
     4441 cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta cgcgcgctca
     4501 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4561 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4621 ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgymctgtag cggcgcatta
     4681 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4741 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4801 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4861 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4921 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4981 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5041 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5101 acgcttacaa tttgtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5161 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5221 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5281 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5341 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5401 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5461 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5521 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5581 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5641 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5701 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5761 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5821 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5881 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5941 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6001 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6061 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6121 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6181 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6241 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6301 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6361 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6421 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6481 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6541 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6601 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6661 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6721 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6781 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6841 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6901 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6961 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7021 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7081 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7141 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7201 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7261 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctgggtacc
     7321 gggccccccc tcgaggtcga cggtatcgat aagcttcacg tgttcgaaac gcggtccgcc
     7381 ctgcagcaac cggaggggga gggagaaggg agcggaaaag tctccaccgg acgcggccat
     7441 ggctcggggg ggggggggca gcggaggagc gcttccggcc gacgtctcgt cgctgattgg
     7501 cttcttttcc tcccgccgtg tgtgaaaaca caaatggcgt gttttggttg gcgtaaggcg
     7561 cctgtcagtt aacggcagcc ggagtgcgca gccgccggca gcctcgctct gcccactggg
     7621 tggggcggga ggtaggtggg gtgaggcgag ctggacgtgc gggcgcggtc ggcctctggc
     7681 ggggcggggg aggggaggga gggtcagcga aagtagctcg cgcgcgagcg gccgcccacc
     7741 ctccccttcc tctgggggag tcgttttacc cgccgccggc cgggcctcgt cgtctgattg
     7801 gctctcgggg cccagaaaac tggcccttgc cattggctcg tgttcgtgca agttgagtcc
     7861 atccgccggc cagcgggggc ggcgaggagg cgctcccagg ttccggccct cccctcggcc
     7921 ccgcgccgca gagtctggcc gcgcgcccct gcgcaacgtg gcaggaagcg cgcgctgggg
     7981 gcggggacgg gcagtagggc tgagcggctg cggggcgggt gcaagcacgt ttccgacttg
     8041 agttgcctca agaggggcgt gctgagccag acctccatcg cgcactccgg ggagtggagg
     8101 gaaggagcga gggctcagtt gggctgtttt ggaggcagga agcacttgct ctcccaaagt
     8161 cgctctgagt tgttatcagt aagggagctg cagtggagta ggcggggaga aggccgcacc
     8221 cttctccgga ggggggaggg gagtgttgca atacctttct gggagttctc tgctgcctcc
     8281 tggcttctga ggaccgccct gggcacatga tggacctgtc gac
//
