LOCUS Exported 8715 bp ds-DNA circular SYN 16-JUN-2022 DEFINITION lentiviral expression of c-Myc. ACCESSION . VERSION . KEYWORDS pCDH-puro-cMyc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8715) AUTHORS Cheng Z, Gong Y, Ma Y, Lu K, Lu X, Pierce LA, Thompson RC, Muller S, Knapp S, Wang J TITLE Inhibition of BET bromodomain targets genetically diverse glioblastoma. JOURNAL Clin Cancer Res. 2013 Apr 1;19(7):1748-59. doi: 10.1158/1078-0432.CCR-12-3066. Epub 2013 Feb 12. PUBMED 23403638 REFERENCE 2 (bases 1 to 8715) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8715 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 180..202 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 194..211 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 195..211 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 226..452 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 453..633 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 680..805 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1298..1531 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1716..1760 /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 2027..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2233..2436 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2387..2407 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2433..2457 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2522..3841 /codon_start=1 /product="human c-Myc proto-oncogene" /label=c-Myc /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA" promoter 3889..4100 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 4113..4381 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" CDS 4408..5007 /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(4408..4427) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4904..4924 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5014..5602 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5067..5087) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5485..5496) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5676..5909 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5981..6102 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6018..6037) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6072..6091 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6142..6277 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6204..6223 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6310..6326) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6310..6326) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6323..6345) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6334..6350 /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)." promoter complement(6358..6388) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6403..6424 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6541..6558) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6712..7300) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6792..6811) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7471..8331) /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 8094..8113 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8332..8436) /gene="bla" /label=AmpR promoter primer_bind 8504..8522 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8560..8582) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8682..8701 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc 61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc 181 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg acgcgtgtag tcttatgcaa 241 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga 301 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa 361 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata 421 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag 481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc 661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc 721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa 781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga 841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc 901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg 961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc 1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac 1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca 1141 agcggccact gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat 1201 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga 1261 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct 1321 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac 1381 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac 1441 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg 1501 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca 1561 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt 1621 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac 1681 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat 1741 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa 1801 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt 1861 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa 1921 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag 1981 acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aattcaaaat tttatcgata ctagtattat 2161 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 2221 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 2281 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 2341 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 2401 aggcgtgtac ggtgggaggt ttatataagc agagctcgtt tagtgaaccg tcagatcgcc 2461 tggagacgcc atccacgctg ttttgacctc catagaagat tctagagcta gcgaattcac 2521 catgcccctc aacgttagct tcaccaacag gaactatgac ctcgactacg actcggtgca 2581 gccgtatttc tactgcgacg aggaggagaa cttctaccag cagcagcagc agagcgagct 2641 gcagcccccg gcgcccagcg aggatatctg gaagaaattc gagctgctgc ccaccccgcc 2701 cctgtcccct agccgccgct ccgggctctg ctcgccctcc tacgttgcgg tcacaccctt 2761 ctcccttcgg ggagacaacg acggcggtgg cgggagcttc tccacggccg accagctgga 2821 gatggtgacc gagctgctgg gaggagacat ggtgaaccag agtttcatct gcgacccgga 2881 cgacgagacc ttcatcaaaa acatcatcat ccaggactgt atgtggagcg gcttctcggc 2941 cgccgccaag ctcgtctcag agaagctggc ctcctaccag gctgcgcgca aagacagcgg 3001 cagcccgaac cccgcccgcg gccacagcgt ctgctccacc tccagcttgt acctgcagga 3061 tctgagcgcc gccgcctcag agtgcatcga cccctcggtg gtcttcccct accctctcaa 3121 cgacagcagc tcgcccaagt cctgcgcctc gcaagactcc agcgccttct ctccgtcctc 3181 ggattctctg ctctcctcga cggagtcctc cccgcagggc agccccgagc ccctggtgct 3241 ccatgaggag acaccgccca ccaccagcag cgactctgag gaggaacaag aagatgagga 3301 agaaatcgat gttgtttctg tggaaaagag gcaggctcct ggcaaaaggt cagagtctgg 3361 atcaccttct gctggaggcc acagcaaacc tcctcacagc ccactggtcc tcaagaggtg 3421 ccacgtctcc acacatcagc acaactacgc agcgcctccc tccactcgga aggactatcc 3481 tgctgccaag agggtcaagt tggacagtgt cagagtcctg agacagatca gcaacaaccg 3541 aaaatgcacc agccccaggt cctcggacac cgaggagaat gtcaagaggc gaacacacaa 3601 cgtcttggag cgccagagga ggaacgagct aaaacggagc ttttttgccc tgcgtgacca 3661 gatcccggag ttggaaaaca atgaaaaggc ccccaaggta gttatcctta aaaaagccac 3721 agcatacatc ctgtccgtcc aagcagagga gcaaaagctc atttctgaag aggacttgtt 3781 gcggaaacga cgagaacagt tgaaacacaa acttgaacag ctacggaact cttgtgcgta 3841 aggatccgcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca 3901 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga 3961 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 4021 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 4081 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc 4141 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct 4201 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg 4261 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc 4321 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat 4381 ccaagctgtg accggcgcct acgctagatg accgagtaca agcccacggt gcgcctcgcc 4441 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc 4501 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa 4561 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc 4621 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc 4681 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc 4741 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg 4801 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc 4861 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac 4921 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg 4981 tgcatgaccc gcaagcccgg tgcctgagtc gacaatcaac ctctggatta caaaatttgt 5041 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct 5101 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat 5161 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg 5221 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag 5281 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc 5341 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg 5401 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc 5461 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc 5521 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc 5581 tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt 5641 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 5701 aagataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 5761 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 5821 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 5881 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 5941 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 6001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 6061 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 6121 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 6181 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 6241 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agaccaaatt 6301 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 6361 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 6421 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 6481 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 6541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 6601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 6661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 6721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 6781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 7021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 7081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 7141 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 7201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 7261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 7321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 7381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 7441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 7501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 7561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 7621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 7681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 7741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 7801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 8041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 8101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 8161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 8221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 8281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 8341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 8401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 8461 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag 8521 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 8581 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 8641 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt 8701 actgagagtg cacca //