LOCUS       sequence-71591-e        7233 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7233)
  AUTHORS   Singh A, Sweeney MF, Yu M, Burger A, Greninger P, Benes C, Haber DA,
            Settleman J
  TITLE     TAK1 Inhibition Promotes Apoptosis in KRAS-Dependent Colon Cancers.
  JOURNAL   Cell. 2012 Feb 17;148(4):639-50.
  PUBMED    22341439
REFERENCE   2  (bases 1 to 7233)
  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: "Cell."; 
            date: "2012-02-17"; volume: "148(4)"; pages: "639-50"
COMMENT     Sequence Label: pLenti6-BMPR1A(Q233D)-V5
FEATURES             Location/Qualifiers
     source          1..7233
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        140..2649
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     protein_bind    2766..2890
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2915..2920,2921..2938,2939..2945)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2999..3679
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(3065..3084)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3999..4304
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4183..4202
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4345..4469)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          4561..4626
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4756..4776
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5201..5335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5238..5257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5292..5311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(5432..5500,5501..6292)
                     /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(5650..5669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6463..7051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6952..6971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7205..7222
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggcaact
       61 cgtgaaaggt aggcggatca gcggccgcag catgcaattc tatattctaa aaacacaaat
      121 gatacttcta aaaaaaaatc atgaatggca tcaactctga atcaaatctt tgcagatgca
      181 cctacttctc atttccactg tcacatcatt tttccagatc tcgctgcctg ttatgtggcc
      241 cacaaaccaa gacacgtttt atggccatta aagctggctg atcgtcgcca aacaccaaat
      301 acataatgaa tatgtacaca ttcgagaaag aagcgatcaa agaagcgtct tcgggcggag
      361 taggagaatg cggaggagaa ggagaacgag ctgatctagt atctctccac aatccaatgc
      421 caactgacca actggccata ttcggagcaa tttgaagcca atttccatcg cctggcgatc
      481 gctccattct tggctatatg tttttcaccg ttacccgggg ccattttcaa agactcgtcg
      541 gcaagataag attgtgtcac tcgctgtctc tcttcatttg tcgaagaatg ctgaggaatt
      601 tcgcgatgac gtcggcgagt attttgaaga atgagaataa tttgtattta tacgaaaatc
      661 agttagtgga attttctaca aaaacatgtt atctatagat aattttgttg caaaatatgt
      721 tgactatgac aaagattgta tgtatatacc tttaatgtat tctcattttc ttatgtattt
      781 ataatggcaa tgatgatact gatgatattt taagatgatg ccagaccaaa aggcttgaat
      841 ttctgcgtct tttgccgaac gcagtgcatg tgcaattgtt gttttttgga atattcaatt
      901 ttcggactgt ccgctttgat ttcagtttct tggcttattc aaaaagcaaa gtaaagccaa
      961 aaaagcgaga tggcaatacc aaatgcggca aaacggtagt ggaaggaaag gggtgcgggg
     1021 cagcggaagg aagggtgggg cggggcgtgg cggggtctgt ggctgggcgc gacgtcaccg
     1081 acgttggagc cactcctttg accatgtgtg cgtgtgtgta ttattcgtgt ctcgccactc
     1141 gccggttgtt tttttctttt tatgctgcgc tctctctagc gccatctcgc ttacgcatgc
     1201 tcaacgcacc gcatgttgcc gtttcctttt atgcgtcatt ttggctcgaa ataggcaatt
     1261 atttaaacaa agattagtca acgaaaacgc taaaataaat aagtctacaa tatggttact
     1321 tattgccatg tgtgtgcagc caacgatagc aacaaaagca acaacacagg tggctttccc
     1381 tctttcactt tttgtttgca agccgcgtgc gagcaagacg gcacgaccgg caaacgcaat
     1441 tacgctgaca aagagcagac gaagttttgg cgaaaaacat caaggcgcct gatacgaatg
     1501 catttgcaat aacaattgcg atatttaata ttgtttatga agctgtttga cttcaaaaca
     1561 cacaaaaaaa aaaataaaac aaattatttg aaagagaatt aggaatcgga cgcttatcgt
     1621 tagggtaaca acaagaaatg cttactgagt cacagcctct ggaaaactgc cgcaagccag
     1681 agagagagag aaaaagaggg agagcagctt agaccgcatg tgcttgtgtg tgaggcgtct
     1741 ctctcttcgt ctctgttgcg caaacgcata gactgcactg agaaaatcga ttacctattt
     1801 tttatgaatg aatatttgca ctattactat tcaaaactat taagatagca atcacattca
     1861 atagccaaat actataccac ctgagcgatg caacgaaatg atcaatttga gcaaaaatgc
     1921 tgcatattta ggacggcatc attatagaaa tgcttcttgc tgtgtacttt tctctcgtct
     1981 ggcagctgtt tcgccgttat tgttaaaacc ggcttaagtt aggtgtgttt tctacgacta
     2041 gtgaatgccc tactagaaga tgtgtgttgc acaaaatgtc cctggaataa ccaatttgaa
     2101 gtgcagatag cagtaaacgt aagctaatat gaatattatt taactgtaat gttttaatat
     2161 cgctggacat tactaataaa cccactataa acacatgtac atatgtatgt tttggcatac
     2221 aatgagtagt tggggaaaaa atgtgtaaaa gcaccgtgac catcacagca taaagataac
     2281 cagctgaagt atcgaatatg agtaaccccc aaattgaatc acatgccgca actgatagga
     2341 cccatggaag tacactcttc atggcgatat acaagacaca cacaagcacg aacacccagt
     2401 tgcggaggaa attctccgta aatgaaaacc caatcggcga acaattcata cccatatatg
     2461 gtaaaagttt tgaacgcgac ttgagagcgg agagcattgc ggctgataag gttttagcgc
     2521 taagcgggct ttataaaacg ggctgcggga ccagttttca tatcactacc gtttgagttc
     2581 ttgtgctgtg tggatactcc tcccgacaca aagccgctcc atcagccagc agtcgtctaa
     2641 tccagagacc ccggatccag atatcgaggc ctgtctagag aagcttgttc gaatctcgag
     2701 tgcgcgcttc cggaggtata cacctaggcg gtaccactgc agtgaattcg gagctctacc
     2761 ggtatacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata
     2821 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     2881 agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2941 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
     3001 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     3061 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     3121 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     3181 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
     3241 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     3301 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     3361 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     3421 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     3481 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     3541 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     3601 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcaggcgg ggcgtaatct
     3661 agaggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
     3721 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga
     3781 agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga
     3841 tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc
     3901 cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat
     3961 gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct
     4021 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
     4081 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtccccc
     4141 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
     4201 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
     4261 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta
     4321 tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat
     4381 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     4441 tctcgttcag ctttcttgta caaagtggtg acgtaagcta gcaggatctt tgtgaaggaa
     4501 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     4561 gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta
     4621 ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc ctttaatgag
     4681 gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac tgctgactct
     4741 caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga ctttccttca
     4801 gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc ttgctttgct
     4861 atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga aaaatattct
     4921 gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt tcttactcca
     4981 cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac ctttagcttt
     5041 ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac tagagatcat
     5101 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5161 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5281 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcccgtt
     5341 taaactacgc gtaattcaaa cagggttctg gcgtcgttct cgtactgttt tccccaggcc
     5401 agtgctttag cgttattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5521 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5581 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5641 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5701 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5821 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5941 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6001 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6301 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6601 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7141 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7201 gcgcagcgag tcagtgagcg aggaagcgga aga
//
