LOCUS Exported 6192 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing JAK2 , part of the Target Accelerator Plasmid Collection. *Please note this is not a true WT JAK2 sequence and contains a V617F mutation.. ACCESSION . VERSION . KEYWORDS pDONR223_JAK2_WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6192) AUTHORS Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn WC TITLE Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles. JOURNAL Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4. PUBMED 27147599 REFERENCE 2 (bases 1 to 6192) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6192 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(4071..4170) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4187..4206) /label=T7 /note="T7 promoter, forward primer" promoter complement(4188..4206) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4211..4227) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4211..4227) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4658..5449 /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 5542..6130 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6031..6050 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agttggcacc atgggaatgg cctgccttac gatgacagaa atggagggaa catccacctc 721 ttctatatat cagaatggtg atatttctgg aaatgccaat tctatgaagc aaatagatcc 781 agttcttcag gtgtatcttt accattccct tgggaaatct gaggcagatt atctgacctt 841 tccatctggg gagtatgttg cagaagaaat ctgtattgct gcttctaaag cttgtggtat 901 cacacctgtg tatcataata tgtttgcttt aatgagtgaa acagaaagga tctggtatcc 961 acccaaccat gtcttccata tagatgagtc aaccaggcat aatgtactct acagaataag 1021 attttacttt cctcgttggt attgcagtgg cagcaacaga gcctatcggc atggaatatc 1081 tcgaggtgct gaagctcctc ttcttgatga ctttgtcatg tcttacctct ttgctcagtg 1141 gcggcatgat tttgtgcacg gatggataaa agtacctgtg actcatgaaa cacaggaaga 1201 atgtcttggg atggcagtgt tagatatgat gagaatagcc aaagaaaacg atcaaacccc 1261 actggccatc tataactcta tcagctacaa gacattctta ccaaaatgta ttcgagcaaa 1321 gatccaagac tatcatattt tgacaaggaa gcgaataagg tacagatttc gcagatttat 1381 tcagcaattc agccaatgca aagccactgc cagaaacttg aaacttaagt atcttataaa 1441 tctggaaact ctgcagtctg ccttctacac agagaaattt gaagtaaaag aacctggaag 1501 tggtccttca ggtgaggaga tttttgcaac cattataata actggaaacg gtggaattca 1561 gtggtcaaga gggaaacata aagaaagtga gacactgaca gaacaggatt tacagttata 1621 ttgcgatttt cctaatatta ttgatgtcag tattaagcaa gcaaaccaag agggttcaaa 1681 tgaaagccga gttgtaacta tccataagca agatggtaaa aatctggaaa ttgaacttag 1741 ctcattaagg gaagctttgt ctttcgtgtc attaattgat ggatattata gattaactgc 1801 agatgcacat cattacctct gtaaagaagt agcacctcca gccgtgcttg aaaatataca 1861 aagcaactgt catggcccaa tttcgatgga ttttgccatt agtaaactga agaaagcagg 1921 taatcagact ggactgtatg tacttcgatg cagtcctaag gactttaata aatatttttt 1981 gacttttgct gtcgagcgag aaaatgtcat tgaatataaa cactgtttga ttacaaaaaa 2041 tgagaatgaa gagtacaacc tcagtgggac aaagaagaac ttcagcagtc ttaaagatct 2101 tttgaattgt taccagatgg aaactgttcg ctcagacaat ataattttcc agtttactaa 2161 atgctgtccc ccaaagccaa aagataaatc aaaccttcta gtcttcagaa cgaatggtgt 2221 ttctgatgta ccaacctcac caacattaca gaggcctact catatgaacc aaatggtgtt 2281 tcacaaaatc agaaatgaag atttgatatt taatgaaagc cttggccaag gcacttttac 2341 aaagattttt aaaggcgtac gaagagaagt aggagactac ggtcaactgc atgaaacaga 2401 agttctttta aaagttctgg ataaagcaca cagaaactat tcagagtctt tctttgaagc 2461 agcaagtatg atgagcaagc tttctcacaa gcatttggtt ttaaattatg gagtatgttt 2521 ctgtggagac gagaatattc tggttcagga gtttgtaaaa tttggatcac tagatacata 2581 tctgaaaaag aataaaaatt gtataaatat attatggaaa cttgaagttg ctaaacagtt 2641 ggcatgggcc atgcattttc tagaagaaaa cacccttatt catgggaatg tatgtgccaa 2701 aaatattctg cttatcagag aagaagacag gaagacagga aatcctcctt tcatcaaact 2761 tagtgatcct ggcattagta ttacagtttt gccaaaggac attcttcagg agagaatacc 2821 atgggtacca cctgaatgca ttgaaaatcc taaaaattta aatttggcaa cagacaaatg 2881 gagttttggt accactttgt gggaaatctg cagtggagga gataaacctc taagtgctct 2941 ggattctcaa agaaagctac aattttatga agataggcat cagcttcctg caccaaagtg 3001 ggcagaatta gcaaacctta taaataattg tatggattat gaaccagatt tcaggccttc 3061 tttcagagcc atcatacgag atcttaacag tttgtttact ccagattatg aactattaac 3121 agaaaatgac atgttaccaa atatgaggat aggtgcccta gggttttctg gtgcctttga 3181 agaccgggat cctacacagt ttgaagagag acatttgaaa tttctacagc aacttggcaa 3241 gggtaatttt gggagtgtgg agatgtgccg gtatgaccct ctacaggaca acactgggga 3301 ggtggtcgct gtaaaaaagc ttcagcatag tactgaagag cacctaagag actttgaaag 3361 ggaaattgaa atcctgaaat ccctacagca tgacaacatt gtaaagtaca agggagtgtg 3421 ctacagtgct ggtcggcgta atctaaaatt aattatggaa tatttaccat atggaagttt 3481 acgagactat cttcaaaaac ataaagaacg gatagatcac ataaaacttc tgcagtacac 3541 atctcagata tgcaagggta tggagtatct tggtacaaaa aggtatatcc acagggatct 3601 ggcaacgaga aatatattgg tggagaacga gaacagagtt aaaattggag attttgggtt 3661 aaccaaagtc ttgccacaag acaaagaata ctataaagta aaagaacctg gtgaaagtcc 3721 catattctgg tatgctccag aatcactgac agagagcaag ttttctgtgg cctcagatgt 3781 ttggagcttt ggagtggttc tgtatgaact tttcacatac attgagaaga gtaaaagtcc 3841 accagcggaa tttatgcgta tgattggcaa tgacaaacaa ggacagatga tcgtgttcca 3901 tttgatagaa cttttgaaga ataatggaag attaccaaga ccagatggat gcccagatga 3961 gatctatatg atcatgacag aatgctggaa caataatgta aatcaacgcc cctcctttag 4021 ggatctagct cttcgagtgg atcaaataag ggataacatg gctggatgat tgccaacttt 4081 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt 4141 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc 4201 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg 4261 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa 4321 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa 4381 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat 4441 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc 4501 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa 4561 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac 4621 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga 4681 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt 4741 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat 4801 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa 4861 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt 4921 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca 4981 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga 5041 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc 5101 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga 5161 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct 5221 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc 5281 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag 5341 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga 5401 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca 5461 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt 5521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 5581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 5641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 5701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 5761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 5821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 5881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 5941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 6001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 6061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 6121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 6181 tgctcacatg tt //