LOCUS       sequence-232063-        6684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6684)
  AUTHORS   Johannessen CM, Boehm JS, Kim SY, Thomas SR, Wardwell L, Johnson LA,
            Emery CM, Stransky N, Cogdill AP, Barretina J, Caponigro G, 
            Hieronymus H, Murray RR, Salehi-Ashtiani K, Hill DE, Vidal M, Zhao 
            JJ, Yang X, Alkan O, Kim S, Harris JL, Wilson CJ, Myer VE, Finan PM,
            Root DE, Roberts TM, Golub T, Flaherty KT, Dummer R, Weber BL, 
            Sellers WR, Schlegel R, Wargo JA, Hahn WC, Garraway LA
  TITLE     COT drives resistance to RAF inhibition through MAP kinase pathway 
            reactivation.
  JOURNAL   Nature. 2010 Dec 16;468(7326):968-72. doi: 10.1038/nature09627
  PUBMED    21107320
REFERENCE   2  (bases 1 to 6684)
  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"; journalName: 
            "Nature"; date: "2010-12-16- 16"; volume: "468"; issue: "7326"; 
            pages: "968-72"
COMMENT     Sequence Label: pDONR223-FLT4
FEATURES             Location/Qualifiers
     source          1..6684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(157..184)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(276..362)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     411..433
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     425..442
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     426..442
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    458..557
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4452..4551)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4568..4587)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4569..4587)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4592..4608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4592..4608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5039..5830
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6665..6682
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
      121 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
      181 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
      241 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
      301 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
      361 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
      421 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
      481 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
      541 gtacaaaaaa gttggcacca tgcagcgggg cgccgcgctg tgcctgcgac tgtggctctg
      601 cctgggactc ctggacggcc tggtgagtgg ctactccatg acccccccga ccttgaacat
      661 cacggaggag tcacacgtca tcgacaccgg tgacagcctg tccatctcct gcaggggaca
      721 gcaccccctc gagtgggctt ggccaggagc tcaggaggcg ccagccaccg gagacaagga
      781 cagcgaggac acgggggtgg tgcgagactg cgagggcaca gacgccaggc cctactgcaa
      841 ggtgttgctg ctgcacgagg tacatgccaa cgacacaggc agctacgtct gctactacaa
      901 gtacatcaag gcacgcatcg agggcaccac ggccgccagc tcctacgtgt tcgtgagaga
      961 ctttgagcag ccattcatca acaagcctga cacgctcttg gtcaacagga aggacgccat
     1021 gtgggtgccc tgtctggtgt ccatccccgg cctcaatgtc acgctgcgct cgcaaagctc
     1081 ggtgctgtgg ccagacgggc aggaggtggt gtgggatgac cggcggggca tgctcgtgtc
     1141 cacgccactg ctgcacgatg ccctgtacct gcagtgcgag accacctggg gagaccagga
     1201 cttcctttcc aaccccttcc tggtgcacat cacaggcaac gagctctatg acatccagct
     1261 gttgcccagg aagtcgctgg agctgctggt aggggagaag ctggtcctga actgcaccgt
     1321 gtgggctgag tttaactcag gtgtcacctt tgactgggac tacccaggga agcaggcaga
     1381 gcggggtaag tgggtgcccg agcgacgctc ccagcagacc cacacagaac tctccagcat
     1441 cctgaccatc cacaacgtca gccagcacga cctgggctcg tatgtgtgca aggccaacaa
     1501 cggcatccag cgatttcggg agagcaccga ggtcattgtg catgaaaatc ccttcatcag
     1561 cgtcgagtgg ctcaaaggac ccatcctgga ggccacggca ggagacgagc tggtgaagct
     1621 gcccgtgaag ctggcagcgt accccccgcc cgagttccag tggtacaagg atggaaaggc
     1681 actgtccggg cgccacagtc cacatgccct ggtgctcaag gaggtgacag aggccagcac
     1741 aggcacctac accctcgccc tgtggaactc cgctgctggc ctgaggcgca acatcagcct
     1801 ggagctggtg gtgaatgtgc ccccccagat acatgagaag gaggcctcct cccccagcat
     1861 ctactcgcgt cacagccgcc aggccctcac ctgcacggcc tacggggtgc ccctgcctct
     1921 cagcatccag tggcactggc ggccctggac accctgcaag atgtttgccc agcgtagtct
     1981 ccggcggcgg cagcagcaag acctcatgcc acagtgccgt gactggaggg cggtgaccac
     2041 gcaggatgcc gtgaacccca tcgagagcct ggacacctgg accgagtttg tggagggaaa
     2101 gaataagact gtgagcaagc tggtgatcca gaatgccaac gtgtctgcca tgtacaagtg
     2161 tgtggtctcc aacaaggtgg gccaggatga gcggctcatc tacttctatg tgaccaccat
     2221 ccccgacggc ttcaccatcg aatccaagcc atccgaggag ctactagagg gccagccggt
     2281 gctcctgagc tgccaagccg acagctacaa gtacgagcat ctgcgctggt accgcctcaa
     2341 cctgtccacg ctgcacgatg cgcacgggaa cccgcttctg ctcgactgca agaacgtgca
     2401 tctgttcgcc acccctctgg ccgccagcct ggaggaggtg gcacctgggg cgcgccacgc
     2461 cacgctcagc ctgagtatcc cccgcgtcgc gcccgagcac gagggccact atgtgtgcga
     2521 agtgcaagac cggcgcagcc atgacaagca ctgccacaag aagtacctgt cggtgcaggc
     2581 cctggaagcc cctcggctca cgcagaactt gaccgacctc ctggtgaacg tgagcgactc
     2641 gctggagatg cagtgcttgg tggccggagc gcacgcgccc agcatcgtgt ggtacaaaga
     2701 cgagaggctg ctggaggaaa agtctggagt cgacttggcg gactccaacc agaagctgag
     2761 catccagcgc gtgcgcgagg aggatgcggg acgctatctg tgcagcgtgt gcaacgccaa
     2821 gggctgcgtc aactcctccg ccagcgtggc cgtggaaggc tccgaggata agggcagcat
     2881 ggagatcgtg atccttgtcg gtaccggcgt catcgctgtc ttcttctggg tcctcctcct
     2941 cctcatcttc tgtaacatga ggaggccggc ccacgcagac atcaagacgg gctacctgtc
     3001 catcatcatg gaccccgggg aggtgcctct ggaggagcaa tgcgaatacc tgtcctacga
     3061 tgccagccag tgggaattcc cccgagagcg gctgcacctg gggagagtgc tcggctacgg
     3121 cgccttcggg aaggtggtgg aagcctccgc tttcggcatc cacaagggca gcagctgtga
     3181 caccgtggcc gtgaaaatgc tgaaagaggg cgccacggcc agcgagcacc gcgcgctgat
     3241 gtcggagctc aagatcctca ttcacatcgg caaccacctc aacgtggtca acctcctcgg
     3301 ggcgtgcacc aagccgcagg gccccctcat ggtgatcgtg gagttctgca agtacggcaa
     3361 cctctccaac ttcctgcgcg ccaagcggga cgccttcagc ccctgcgcgg agaagtctcc
     3421 cgagcagcgc ggacgcttcc gcgccatggt ggagctcgcc aggctggatc ggaggcggcc
     3481 ggggagcagc gacagggtcc tcttcgcgcg gttctcgaag accgagggcg gagcgaggcg
     3541 ggcttctcca gaccaagaag ctgaggacct gtggctgagc ccgctgacca tggaagatct
     3601 tgtctgctac agcttccagg tggccagagg gatggagttc ctggcttccc gaaagtgcat
     3661 ccacagagac ctggctgctc ggaacattct gctgtcggaa agcgacgtgg tgaagatctg
     3721 tgactttggc cttgcccggg acatctacaa agaccccgac tacgtccgca agggcagtgc
     3781 ccggctgccc ctgaagtgga tggcccctga aagcatcttc gacaaggtgt acaccacgca
     3841 gagtgacgtg tggtcctttg gggtgcttct ctgggagatc ttctctctgg gggcctcccc
     3901 gtaccctggg gtgcagatca atgaggagtt ctgccagcgg ctgagagacg gcacaaggat
     3961 gagggccccg gagctggcca ctcccgccat acgccgcatc atgctgaact gctggtccgg
     4021 agaccccaag gcgagacctg cattctcgga gctggtggag atcctggggg acctgctcca
     4081 gggcaggggc ctgcaagagg aagaggaggt ctgcatggcc ccgcgcagct ctcagagctc
     4141 agaagagggc agcttctcgc aggtgtccac catggcccta cacatcgccc aggctgacgc
     4201 tgaggacagc ccgccaagcc tgcagcgcca cagcctggcc gccaggtatt acaactgggt
     4261 gtcctttccc gggtgcctgg ccagaggggc tgagacccgt ggttcctcca ggatgaagac
     4321 atttgaggaa ttccccatga ccccaacgac ctacaaaggc tctgtggaca accagacaga
     4381 cagtgggatg gtgctggcct cggaggagtt tgagcagata gagagcaggc atagacaaga
     4441 aagcggcttc aggccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     4501 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4561 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     4621 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
     4681 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
     4741 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
     4801 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     4861 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
     4921 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     4981 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     5041 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     5101 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     5161 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     5221 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     5281 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     5341 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     5401 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     5461 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     5521 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     5581 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     5641 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     5701 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     5761 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     5821 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6541 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6601 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6661 gcgcagcgag tcagtgagcg agga
//
