LOCUS Exported 7585 bp ds-DNA circular SYN 20-DEC-2021 DEFINITION Empty vector to express overexpression cDNA. ACCESSION . VERSION . KEYWORDS pHAGE_puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7585) AUTHORS Lu B, Klingbeil O, Tarumoto Y, Somerville TDD, Huang YH, Wei Y, Wai DC, Low JKK, Milazzo JP, Wu XS, Cao Z, Yan X, Demerdash OE, Huang G, Mackay JP, Kinney JB, Shi J, Vakoc CR TITLE A Transcription Factor Addiction in Leukemia Imposed by the MLL Promoter Sequence. JOURNAL Cancer Cell. 2018 Dec 10;34(6):970-981.e8. doi: 10.1016/j.ccell.2018.10.015. Epub 2018 Nov 29. PUBMED 30503706 REFERENCE 2 (bases 1 to 7585) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 20, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7585 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 52..71 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 140..739 /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(140..159) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 636..656 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 753..1341 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(806..826) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1224..1235) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 1416..1649 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(1768..1786) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1854..1958 /gene="bla" /label=AmpR promoter CDS 1959..2819 /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(2177..2196) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2990..3578 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3479..3498 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3732..3749 /label=L4440 /note="L4440 vector, forward primer" primer_bind 3841..3860 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 3989..4010 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4025..4055 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4063..4079 /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 4068..4090 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4087..4103 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4087..4103 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 4153..4172 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 4364..4997 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 5044..5169 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 5666..5899 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6084..6128 /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 6421..6538 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 6590..6893 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6894..7097 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7048..7068 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 7094..7118 /label=LNCX /note="Human CMV promoter, forward primer" promoter 7205..119 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(7240..7263) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" ORIGIN 1 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca 61 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga 121 cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 181 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 241 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 301 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 361 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 421 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 481 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 541 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 601 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 661 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 721 ccgcaagccc ggtgcctgaa tcgatagatc ctaatcaacc tctggattac aaaatttgtg 781 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 841 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 901 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 961 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 1021 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 1081 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 1141 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 1201 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 1261 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 1321 ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca aggcagctgt 1381 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 1441 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 1501 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 1561 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 1621 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 1681 tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa ttaaggaaag 1741 ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg 1801 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 1861 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 1921 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 1981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 2101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 2161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc 2221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 2281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 2341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 2401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 2461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 2521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 2581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 2641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 2701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 2761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 2821 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 2881 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 2941 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 3001 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 3061 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 3121 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 3181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 3241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 3301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 3361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 3421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 3481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 3541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 3601 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 3661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 3721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 3781 ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac taattttttt 3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 3901 cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg tttcccgact 3961 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc 4021 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat 4081 ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca tttttttcac 4141 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact 4201 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg 4261 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt 4321 aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg gctaattcac 4381 tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct 4441 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc 4501 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc 4561 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag 4621 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac 4681 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg 4741 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct 4801 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 4861 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 4921 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 4981 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag 5041 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg 5101 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc 5161 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg 5221 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa 5281 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga 5341 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta 5401 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac 5461 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga 5521 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata 5581 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc 5641 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag 5701 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg 5761 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc 5821 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc 5881 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg 5941 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga 6001 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg 6061 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg 6121 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa 6181 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata 6241 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac 6301 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc 6361 gattagtgaa cggatctcga cggtatcgcc gaattcacaa atggcagtat tcatccacaa 6421 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat 6481 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg 6541 ggtttattac agggacagca gagatccagt ttggactagt ggagttccgc gttacataac 6601 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 6661 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 6721 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 6781 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 6841 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 6901 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 6961 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 7021 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 7081 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct 7141 gttttgacct ccatagaaga caccggcggc cgcaccggtc tgcagctagc tcgagaattc 7201 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg 7261 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt 7321 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta 7381 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta 7441 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc 7501 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga 7561 aggggtgggt ccgggggcgg gctca //