LOCUS       sequence-227371-        9858 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2001 Mar . 7(3):673-82."
COMMENT     Sequence Label: pCEP4-HA-BAX
FEATURES             Location/Qualifiers
     source          1..9858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             656..682
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     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             join(5840..5908,5909..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
//
