LOCUS Exported 9858 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCEP4-HA-BAX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9858) AUTHORS Yu J, Zhang L, Hwang PM, Kinzler KW, Vogelstein B TITLE PUMA induces the rapid apoptosis of colorectal cancer cells. JOURNAL Mol Cell. 2001 Mar . 7(3):673-82. PUBMED 11463391 REFERENCE 2 (bases 1 to 9858) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9858 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 581..605 /label=LNCX /note="Human CMV promoter, forward primer" polyA_signal 1286..1420 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1310..1329) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1364..1383 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1563..1585) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5649..5667) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5735..5839 /gene="bla" /label=AmpR promoter CDS 5840..6700 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6058..6077) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6871..7459 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7360..7379 /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 7890..8035 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" CDS 8078..9115 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE MGEAN" primer_bind complement(9113..9132) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 9157..9205 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 601 agatctctag aagctgggta ccgaaatggg atacgatgtt ccagattacg cttcgtacga 661 tgttccagat tacgcaagct tgatggacgg gtccggggag cagcccagag gcggggggcc 721 caccagctct gagcagatca tgaagacagg ggcccttttg cttcagggtt tcatccagga 781 tcgagcaggg cgaatggggg gggaggcacc cgagctggcc ctggacccgg tgcctcagga 841 tgcgtccacc aagaagctga gcgagtgtct caagcgcatc ggggacgaac tggacagtaa 901 catggagctg cagaggatga ttgccgccgt ggacacagac tccccccgag aggtcttttt 961 ccgagtggca gctgacatgt tttctgacgg caacttcaac tggggccggg ttgtcgccct 1021 tttctacttt gccagcaaac tggtgctcaa ggccctgtgc accaaggtgc cggaactgat 1081 cagaaccatc atgggctgga cattggactt cctccgggag cggctgttgg gctggatcca 1141 agaccagggt ggttgggacg gcctcctctc ctactttggg acgcccacgt ggcagaccgt 1201 gaccatcttt gtggcgggag tgctcaccgc ctcactcacc atctggaaga agatgggctg 1261 ataagtcgag gggccggcaa ggccggatcc agacatgata agatacattg atgagtttgg 1321 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat 1381 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca 1441 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta 1501 caaatgtggt atggctgatt atgatccggc tgcctcgcgc gtttcggtga tgacggtgaa 1561 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg 1621 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg 1681 aggtcgactc tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc 1741 tgccccttct tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa 1801 ctgggccctg ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg 1861 tagttgacat ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg 1921 gagcagactt tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc 1981 tgaagctctt acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg 2041 gaggctacac caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag 2101 agggcattag caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc 2161 ttcccgggta gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca 2221 acgggaagca tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc 2281 tcccacccca tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt 2341 gaaccatttt agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct 2401 gaatcttcgc ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt 2461 gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata 2521 aaatttggac ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa 2581 accccttggg caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag 2641 aaatccatgg ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg 2701 ctatgggcaa cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag 2761 gcagggacca agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga 2821 gtggacgccg acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg 2881 ggctccacgc caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg 2941 tggagtgggg gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa 3001 gggtgtaata acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca 3061 caaaaccact aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag 3121 gggcgcgatt gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca 3181 cagggttgtt ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca 3241 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg 3301 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg 3361 tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg 3421 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg 3481 tagcatatgc tatcctcatg catatacagt cagcatatga tacccagtag tagagtggga 3541 gtgctatcct ttgcatatgc cgccacctcc caagggggcg tgaattttcg ctgcttgtcc 3601 ttttcctgct ggttgctccc attcttaggt gaatttaagg aggccaggct aaagccgtcg 3661 catgtctgat tgctcaccag gtaaatgtcg ctaatgtttt ccaacgcgag aaggtgttga 3721 gcgcggagct gagtgacgtg acaacatggg tatgcccaat tgccccatgt tgggaggacg 3781 aaaatggtga caagacagat ggccagaaat acaccaacag cacgcatgat gtctactggg 3841 gatttattct ttagtgcggg ggaatacacg gcttttaata cgattgaggg cgtctcctaa 3901 caagttacat cactcctgcc cttcctcacc ctcatctcca tcacctcctt catctccgtc 3961 atctccgtca tcaccctccg cggcagcccc ttccaccata ggtggaaacc agggaggcaa 4021 atctactcca tcgtcaaagc tgcacacagt caccctgata ttgcaggtag gagcgggctt 4081 tgtcataaca aggtccttaa tcgcatcctt caaaacctca gcaaatatat gagtttgtaa 4141 aaagaccatg aaataacaga caatggactc ccttagcggg ccaggttgtg ggccgggtcc 4201 aggggccatt ccaaagggga gacgactcaa tggtgtaaga cgacattgtg gaatagcaag 4261 ggcagttcct cgccttaggt tgtaaaggga ggtcttacta cctccatata cgaacacacc 4321 ggcgacccaa gttccttcgt cggtagtcct ttctacgtga ctcctagcca ggagagctct 4381 taaaccttct gcaatgttct caaatttcgg gttggaacct ccttgaccac gatgctttcc 4441 aaaccaccct ccttttttgc gcctgcctcc atcaccctga ccccggggtc cagtgcttgg 4501 gccttctcct gggtcatctg cggggccctg ctctatcgct cccgggggca cgtcaggctc 4561 accatctggg ccaccttctt ggtggtattc aaaataatcg gcttccccta cagggtggaa 4621 aaatggcctt ctacctggag ggggcctgcg cggtggagac ccggatgatg atgactgact 4681 actgggactc ctgggcctct tttctccacg tccacgacct ctccccctgg ctctttcacg 4741 acttcccccc ctggctcttt cacgtcctct accccggcgg cctccactac ctcctcgacc 4801 ccggcctcca ctacctcctc gaccccggcc tccactgcct cctcgacccc ggcctccacc 4861 tcctgctcct gccnnncctc ctgctcctgc ccctcctcct gctcctgccc ctcctgctcc 4921 tgcccctccc gctcctgctc ctgctcctgt tccaccgtgg gtccctttgc agccaatgca 4981 acttggacgt ttttggggtc tccggacacc atctctatgt cttggccctg atcctgagcc 5041 gcccggggct cctggtcttc cgcctcctcg tcctcgtcct cttccccgtc ctcgtccatg 5101 gttatcaccc cctcttcttt gaggtccact gccgccggag ccttctggtc cagatgtgtc 5161 tcccttctct cctaggccat ttccaggtcc tgtacctggc ccctcgtcag acatgattca 5221 cactaaaaga gatcaataga catctttatt agacgacgct cagtgaatac agggagtgca 5281 gactcctgcc ccctccaaca gcccccccac cctcatcccc ttcatggtcg ctgtcagaca 5341 gatccaggtc tgaaaattcc ccatcctccg aaccatcctc gtcctcatca ccaattactc 5401 gcagcccgga aaactcccgc tgaacatcct caagatttgc gtcctgagcc tcaagccagg 5461 cctcaaattc ctcgtccccc tttttgctgg acggtaggga tggggattct cgggacccct 5521 cctcttcctc ttcaaggtca ccagacagag atgctactgg ggcaacggaa gaaaagctgg 5581 gtgcggcctg tgaggatcag cttatcgatg ataagctgtc aaacatgaga attcttgaag 5641 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc 5701 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 5761 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 5821 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 5881 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 5941 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 6001 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 6061 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca 6121 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 6181 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 6241 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 6301 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 6361 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg 6421 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 6481 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 6541 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 6601 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 6661 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 6721 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 6781 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 6841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 6901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 6961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct 7021 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 7081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 7141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 7201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 7261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 7321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 7381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 7441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 7501 ctggccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa 7561 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc 7621 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccccgagat gcgccgcgtg 7681 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc 7741 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg 7801 tgccgccctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag 7861 aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat 7921 tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tccgaggtcc acttcgcata 7981 ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaacagcg 8041 tcaacagcgt gccgcagatc ccggggggca atgagatatg aaaaagcctg aactcaccgc 8101 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct 8161 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct 8221 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc 8281 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac 8341 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact 8401 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag 8461 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg 8521 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga 8581 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg 8641 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa 8701 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga 8761 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta 8821 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg 8881 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg 8941 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca 9001 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag 9061 tggaaaccga cgccccagca ctcgtccgga tcgggagatg ggggaggcta actgaaacac 9121 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 9181 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 9241 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 9301 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 9361 cctgccatag ccactggccc cgtgggttag ggacggggtc ccccatgggg aatggtttat 9421 ggttcgtggg ggttattatt ttgggcgttg cgtggggtca ggtccacgac tggactgagc 9481 agacagaccc atggtttttg gatggcctgg gcatggaccg catgtactgg cgcgacacga 9541 acaccgggcg tctgtggctg ccaaacaccc ccgaccccca aaaaccaccg cgcggatttc 9601 tggcgtgcca agctagtcga ccaattctca tgtttgacag cttatcatcg cagatccggg 9661 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga 9721 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct 9781 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc 9841 caatatgacc gccatgtt //