LOCUS       sequence-386373-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CRKL Y207F + W275L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  AUTHORS   Cheung HW, Du J, Boehm JS, He F, Weir BA, Wang X, Butaney M, Sequist
            LV, Luo B, Engelman JA, Root DE, Meyerson M, Golub TR, Janne PA, 
            Hahn WC
  TITLE     Amplification of CRKL induces transformation and epidermal growth 
            factor receptor inhibitor resistance in human non-small cell lung 
            cancers.
  JOURNAL   Cancer Discov. 2011 Dec;1(7):608-25. doi: 
            10.1158/2159-8290.CD-11-0046. Epub 2011 Oct 17.
  PUBMED    22586683
REFERENCE   2  (bases 1 to 6011)
  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: 
            "10.1158/2159-8290.CD-11-0046"; journalName: "Cancer Discov"; date: 
            "2011-12"; volume: "1"; issue: "7"; pages: "608-25"
COMMENT     Sequence Label: pWzl_CRKL-Y207F+W275L
FEATURES             Location/Qualifiers
     source          1..6011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(982..1773,1774..1842))
                     /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     1605..1624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1843..1947)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1978..2447
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2512..2711
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2912..3328
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3258..3280
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3296..3312
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    3383..3407
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4323..4347)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4398..4859
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4454..4471)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4681..4700
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             5475..5944
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata
      961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg
     1981 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2041 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2101 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     2161 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     2221 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     2281 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     2341 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     2401 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     2461 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg caagctggcc
     2521 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     2581 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     2641 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
     2701 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     2761 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     2821 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     2881 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     2941 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     3001 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     3061 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     3121 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     3181 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     3241 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     3301 tccagccctc actccttctc taggcgccgg ccggatccag aattcactcg agatacgtaa
     3361 aacaattgaa gatctagtcg agacaagttt gtacaaaaaa gttggcacca tgtcctccgc
     3421 caggttcgac tcctcggacc gctccgcctg gtatatgggg ccggtgtctc gccaggaggc
     3481 gcagacccgg ctccagggcc agcgccacgg tatgttcctc gtccgcgatt cttccacctg
     3541 ccctggggac tatgtgctgt cggtgtccga gaactcgcgg gtctcccact acatcatcaa
     3601 ctcgctgccc aaccgccgtt ttaagatcgg ggaccaggaa tttgaccatt tgccggccct
     3661 gctggagttt tacaagatcc actacctgga caccaccacc ctcatcgagc ctgcgcccag
     3721 gtatccaagc ccaccaatgg gatctgtctc agcacccaac ctgcctacag cagaagataa
     3781 cctggaatat gtacggactc tgtatgattt tcctgggaat gatgccgaag acctgccctt
     3841 taaaaagggt gagatcctag tgataataga gaagcctgaa gaacagtggt ggagtgcccg
     3901 gaacaaggat ggccgggttg ggatgattcc tgtcccttat gtcgaaaagc ttgtgagatc
     3961 ctcaccacac ggaaagcatg gaaataggaa ttccaacagt tatgggatcc cagaacctgc
     4021 tcatgcattc gctcaacctc agaccacaac tcctctacct gcagtttccg gttctcctgg
     4081 ggcagcaatc acccctttgc catccacaca gaatggacct gtctttgcga aagcaatcca
     4141 gaaaagagta ccctgtgctt atgacaagac tgccttggca ttagaggttg gtgacatcgt
     4201 gaaagtcaca aggatgaata taaatggcca gttggaaggc gaagtgaacg ggcgcaaagg
     4261 gcttttcccc tttacgcacg tcaaaatctt tgaccctcaa aacccagatg aaaacgagtg
     4321 attgccaact ttcttgtaca aacttgtctc gaccaattcc ggttattttc caccatattg
     4381 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     4441 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     4501 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     4561 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     4621 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     4681 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     4741 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     4801 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     4861 accatgaaaa catttaacat ttctcaacag gatctagaat tagtagaagt agcgacagag
     4921 aagattacaa tgctttatga ggataataaa catcatgtgg gagcggcaat tcgtacgaaa
     4981 acaggagaaa tcatttcggc agtacatatt gaagcgtata taggacgagt aactgtttgt
     5041 gcagaagcca ttgcgattgg tagtgcagtt tcgaatggac aaaaggattt tgacacgatt
     5101 gtagctgtta gacaccctta ttctgacgaa gtagatagaa gtattcgagt ggtaagtcct
     5161 tgtggtatgt gtagggagtt gatttcagac tatgcaccag attgttttgt gttaatagaa
     5221 atgaatggca agttagtcaa aactacgatt gaagaactca ttccactcaa atatacccga
     5281 aattaaaagt tttaccatac caagctcatc gataaaataa aagattttat ttagtctcca
     5341 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5401 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     5461 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5521 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     5581 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     5641 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     5701 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5761 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     5821 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5881 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5941 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     6001 ggccatagtt g
//
