LOCUS Exported 9155 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of SRSF2 P95H. ACCESSION . VERSION . KEYWORDS pHAGE-SRSF2-P95H SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9155) AUTHORS Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z, Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott KL, Yi S, Sahni N, Liang H, Mills GB TITLE Systematic Functional Annotation of Somatic Mutations in Cancer. JOURNAL Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 10.1016/j.ccell.2018.01.021. PUBMED 29533785 REFERENCE 2 (bases 1 to 9155) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9155 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2058..2175 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2223..3401 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2454..3392 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3349..3369 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 3421..3445 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(4115..4139) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter 4147..4165 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4190..4209) /label=T7 /note="T7 promoter, forward primer" promoter complement(4191..4209) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind complement(4215..4235) /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" CDS 4289..4330 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 4351..4918 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 4368..4920 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4512..4529) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4739..4758 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4932..5531 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4932..4951) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5428..5448 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5545..6133 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5598..5618) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6016..6027) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6208..6441 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6560..6578) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6646..6750 /gene="bla" /label=AmpR promoter CDS 6751..7611 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6969..6988) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7782..8370 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8271..8290 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8524..8541 /label=L4440 /note="L4440 vector, forward primer" primer_bind 8633..8652 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 8781..8802 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8817..8847 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8855..8871 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 8860..8882 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8879..8895 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8879..8895 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8945..8964 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt 2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa 2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg 2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa 2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt 2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct 2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg 2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct 2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg 2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc 2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc 2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac 3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga 3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc 3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg 3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga 3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt 3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga 3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccct gcagatatca 3421 acaagtttgt acaaaaaagc aggcaccatg agctacggcc gcccccctcc cgatgtggag 3481 ggtatgacct ccctcaaggt ggacaacctg acctaccgca cctcgcccga cacgctgagg 3541 cgcgtcttcg agaagtacgg gcgcgtcggc gacgtgtaca tcccgcggga tcgctacacc 3601 aaggagtccc gcggcttcgc cttcgttcgc tttcacgaca agcgcgacgc tgaggacgct 3661 atggatgcca tggacggggc cgtgctggac ggccgcgagc tgcgggtgca aatggcgcgc 3721 tacggccgcc acccggactc acaccacagc cgccggggac cgccaccccg caggtacggg 3781 ggcggtggct acggacgccg gagccgcagc cctaggcggc gtcgccgcag ccgatcccgg 3841 agtcggagcc gttccaggtc tcgcagccga tctcgctaca gccgctcgaa gtctcggtcc 3901 cgcactcgtt ctcgatctcg gtcgacctcc aagtccagat ccgcacgaag gtccaagtcc 3961 aagtcctcgt cggtctccag atctcgttcg cggtccaggt cccggtctcg gtccaggagt 4021 cctcccccag tgtccaagag ggaatccaaa tccaggtcgc gatcgaagag tccccccaag 4081 tctcctgaag aggaaggagc ggtgtcctct tagaacccag ctttcttgta caaagtggtg 4141 atatccaatt aaccctcact aaaggtatta tgagaaagtt gaatagtagc cctatagtga 4201 gtcgtattac cacccaactt ttctatacaa agtggttgat atccagcaca gtggcggccg 4261 ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct 4321 cgattctacg cgtaccggtt agtaatgaga tccctccccc ccccctaacg ttactggccg 4381 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc 4441 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag 4501 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt 4561 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa 4621 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc 4681 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg 4741 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat 4801 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa 4861 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata 4921 tggccacaca tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 4981 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 5041 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 5101 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 5161 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 5221 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 5281 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 5341 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 5401 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 5461 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 5521 ccggtgccta aatcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg 5581 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5641 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5701 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5761 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5821 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5881 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5941 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6001 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6061 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6121 gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta 6181 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag 6241 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 6301 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6361 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6421 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6481 ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat 6541 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 6601 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 6661 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 6721 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 6781 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 6841 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 6901 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 6961 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt 7021 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 7081 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 7141 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 7201 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 7261 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 7321 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 7381 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 7441 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 7501 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 7561 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 7621 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 7681 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 7741 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 7801 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 7861 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 7921 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 7981 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 8041 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 8101 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 8161 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 8221 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 8281 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 8341 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 8401 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 8461 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 8521 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 8581 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg 8641 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 8701 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg 8761 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta 8821 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca 8881 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct 8941 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact 9001 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac 9061 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat 9121 tgtatttgtt aaatatgtac tacaaactta gtagt //