LOCUS Exported 9328 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral expression of MCM5__LYN fusion. ACCESSION . VERSION . KEYWORDS pHAGE-MCM5__LYN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9328) 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 9328) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9328 /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(3868..3892) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(4388..4408) /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" CDS 4462..4503 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 4524..5091 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 4541..5093 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4685..4702) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4912..4931 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5105..5704 /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(5105..5124) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5601..5621 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5718..6306 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5771..5791) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6189..6200) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6381..6614 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6733..6751) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6819..6923 /gene="bla" /label=AmpR promoter CDS 6924..7784 /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(7142..7161) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7955..8543 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8444..8463 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8697..8714 /label=L4440 /note="L4440 vector, forward primer" primer_bind 8806..8825 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 8954..8975 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8990..9020 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9028..9044 /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 9033..9055 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9052..9068 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9052..9068 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9118..9137 /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 acaaaaaagt tggcatgtcg ggattcgacg atcctggcat tttctacagc 3481 gacagcttcg ggggcgacgc ccaggccgac gaggggcagg cccgcaaatc gcagctgcag 3541 aggcgcttca aggagttcct gcggcagtac cgagtgggca ccgaccgcac gggcttcacc 3601 ttcaaataca gggatgaact caagcggcat tacaacctgg gggagtactg gattgaggtg 3661 gagatggagg atctggccag ctttgatgag gacctggccg actacttgta caagcagcca 3721 gccgagcacc tgcagctgct ggaggaagct gccaaggagg tagctgatga ggtgacccgg 3781 ccccggcctt ctggggagga ggtgctccag gacatccagg tcatgctcaa gtcggacgcc 3841 agcccttcca gcattcgtag cctgaagtcc cagctttctt gtacaaagtg gttcattgca 3901 gagggaatgg catacatcga gcggaagaac tacattcacc gggacctgcg agcagctaat 3961 gttctggtct ccgagtcact catgtgcaaa attgcagatt ttggccttgc tagagtaatt 4021 gaagataatg agtacacagc aagggaaggt gctaagttcc ctattaagtg gacggctcca 4081 gaagcaatca actttggatg tttcactatt aagtctgatg tgtggtcctt tggaatcctc 4141 ctatacgaaa ttgtcaccta tgggaaaatt ccctacccag ggagaactaa tgccgacgtg 4201 atgaccgccc tgtcccaggg ctacaggatg ccccgtgtgg agaactgccc agatgagctc 4261 tatgacatta tgaaaatgtg ctggaaagaa aaggcagaag agagaccaac gtttgactac 4321 ttacagagcg tcctggatga tttctacaca gccacggaag ggcaatacca gcagcagcct 4381 tagcacccaa cttttctata caaagtggtt gatatccagc acagtggcgg ccgctcgagt 4441 ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct 4501 acgcgtaccg gttagtaatg agatccctcc ccccccccta acgttactgg ccgaagccgc 4561 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt 4621 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt 4681 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg 4741 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca 4801 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg 4861 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc 4921 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct 4981 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta 5041 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac 5101 acatatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc 5161 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc 5221 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 5281 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 5341 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 5401 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 5461 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 5521 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 5581 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 5641 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 5701 ctaaatcgat agatcctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta 5761 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 5821 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 5881 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 5941 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 6001 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 6061 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt 6121 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 6181 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 6241 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 6301 ccccgcctga gatcctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6361 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct 6421 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6481 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 6541 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6601 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6661 gaaatgaata tcagagagtg agaggcccgg gttaattaag gaaagggcta gatcattctt 6721 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg 6781 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 6841 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 6901 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct 6961 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag 7021 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta 7081 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc 7141 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca 7201 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg 7261 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg 7321 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca 7381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa 7441 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa 7501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata 7561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat 7621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc 7681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata 7741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt 7801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 7861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 7921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 7981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 8041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 8101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 8161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 8221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 8281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 8341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 8401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 8461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 8521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 8581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 8641 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg 8701 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt 8761 ggccgattca ttaatgcagc aagctcatgg ctgactaatt ttttttattt atgcagaggc 8821 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 8881 aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa gcgggcagtg 8941 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 9001 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 9061 gctatgacat gattacgaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 9121 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta 9181 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt 9241 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt 9301 gttaaatatg tactacaaac ttagtagt //