LOCUS       sequence-200263-        4381 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cre-recombinase, a barcoded HDR template to introduce a 
            G13A mutation into the endogenous Kras locus, and a Kras-targeting 
            sgRNA to enhance HDR. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4381)
  AUTHORS   Winters IP, Chiou SH, Paulk NK, McFarland CD, Lalgudi PV, Ma RK, 
            Lisowski L, Connolly AJ, Petrov DA, Kay MA, Winslow MM
  TITLE     Multiplexed in vivo homology-directed repair and tumor barcoding 
            enables parallel quantification of Kras variant oncogenicity.
  JOURNAL   Nat Commun. 2017 Dec 12;8(1):2053. doi: 10.1038/s41467-017-01519-y.
  PUBMED    29233960
REFERENCE   2  (bases 1 to 4381)
  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: "Nat 
            Commun."; date: "2017-12-12"; volume: "8(1):2053. doi"; pages: " 
            10.1038/s41467-017-01519-y"
COMMENT     Sequence Label: AAV-KrasG13A/sgKras/Cre
FEATURES             Location/Qualifiers
     source          1..4381
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      35..44
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2069..2317
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2069..2089
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2240..2259
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2338..2413
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2466..2965
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2501..2524)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2898..2917
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2969..2989
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2990..4018
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
ORIGIN
        1 ctgatatcat cgatcgcgat gcattaatta agcggccgcc atggcagttc ttttgtatcc
       61 atttgtctct ttatctgcaa taatatatgt tctacaaggg aaaggagata aagtgtatat
      121 attaatttta tttcaaaaat gtttggaaaa gcagactagg aaacaacagt gttgtattca
      181 gggctgttaa ctgtatacag tgccagcact tagtgcacct gttgggagtg cagtgtatta
      241 taagcaaggc agaagtcaca gaggttactg agaaactctt ttatgttctt tacatttttt
      301 ttcttgccaa aaatagtctt tatctctgaa tttcattcaa ttttatttcc ttgtacagag
      361 gtactgtttt tagtactcaa aggattatat gttaaaattg atctgatcct ataccaaata
      421 tttaaaccca tttcccacag atagtttata gattaaaatg ccatcaaata ttatctttat
      481 ttgcttcatt gcctaaatat gtctttcccc agcacagtgc agttttgaca ccagcttcgg
      541 cttcctattt tgttgccctt tattgcagaa ctgctctgat ggctttaaaa cagttctctg
      601 cagttgttgg ctccaacaca gatgttctta ggctacctaa cttctaactt ttaatatcca
      661 gtcaacaaag aataccgcaa gggtaggtgt tgggatagct gtcgacaagc tcatgcgggt
      721 gtgtccacag ggtatagcgt actatgcaga atatttgtac tgagtgaagt catgatacat
      781 tcctttgaga gccattagct gctacaaaac agtaatctgg ctgtttagat caacaagcta
      841 aatgatagaa gatgaaagta ctggtttcca tgtattttta ttaagtgttg atgagaaagt
      901 tgtaagtgac ttacaggtta ctctgtacat ctgtagtcac tgaattcgga atatcttaga
      961 gttttacaca cctaggtgag tgttaaaata ttgataaagt ttttgataat cttgtgtgag
     1021 acatgttcta atttagttgt attttattat ttttattgta aggcctgctg aaaatgactg
     1081 agtataaact agtagtcgtt ggagctggtg ccgtggggaa atcagcgcta acaatacagc
     1141 taattcagaa tcactttgtg gatgagtatg accctacgat agaggtaacg ctgctctaca
     1201 gtctgcgtac gcttgtaaag gacggcagcc agccgctttg aaaaagatat catttttata
     1261 tttattagaa aattatattg aaagttattt cagttatatg tgatgtcctt tagttccaag
     1321 gctttaaact gggtgttagg gaaccatagg tgcaagaaag tccacttctc atgagagctc
     1381 accacagaga aagaaagtcc acttctcatg agagcttacc acagagagta aggtccaaga
     1441 tgccatccgt gctttgtatt tgcattttga taaataaatt tcatgatttt gtgagagtta
     1501 agaatatagc tttgcctttg atctattcct gaaaggtttg ttgttgtttt atgggtttgt
     1561 tgttattgtg ttgttgtttg gtgtctttta ggcagggtct tactatgcag ccctgaccgg
     1621 cctagaactc aactatgtag accagactag ctctgaattc tcagagatcc atctacctgc
     1681 ctctgctgcc cagatgctgc gattaaagat gtacaccacc atgtacagtg aaagttactc
     1741 ttttgaaaac actttattgt tatagattac ttggaatgtt gatgtgatga gaaaatttga
     1801 ttatactagg aaaatagaat cacttgatgg ttgagctttt tttttaagac ttatttattt
     1861 tatgtaagtg agtacactgc ttctctcttc agacacacca gaagagggca tcagatccca
     1921 ttaagatggc tgtgagccac catgtggttg ctgggaattg aactcaggac ctctggaaag
     1981 gcagtcagtg ctcttttttt gtttaaagat ttatttattt ttatgaatac agtgtcactg
     2041 tcttcataca cacaagagct cgagctgtga gggcctattt cccatgattc cttcatattt
     2101 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     2161 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     2221 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     2281 cttggcttta tatatcttgt ggaaaggacc aaacaccgac tgagtataaa cttgtgggtt
     2341 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     2401 accgagtcgg tgcttttttg tttggcgcgc ctatggctta attaacatgg cggccgcatt
     2461 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     2521 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     2581 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     2641 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     2701 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     2761 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     2821 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     2881 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     2941 ctcttcctca tctccgggac ccgggatgcc caagaagaag aggaaggtgt ccaatttact
     3001 gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa
     3061 gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct
     3121 tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc
     3181 cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt
     3241 aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc
     3301 acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt
     3361 tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt
     3421 tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg
     3481 gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat
     3541 ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag
     3601 caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat
     3661 ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa
     3721 tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt
     3781 tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc
     3841 ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc
     3901 aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca tgaactatat
     3961 ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattagcc
     4021 atgtttaagg gttcctctag tgtacacgcg ttgtgccttc tagttgccag ccatctgttg
     4081 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4141 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4201 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaac
     4261 gcgtgatatc agatctgtta cgtagataag tagcatggcg ggttaatcat taactacaag
     4321 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4381 g
//
