LOCUS       sequence-198847-       10024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10024)
  AUTHORS   Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B
  TITLE     A model for p53-induced apoptosis.
  JOURNAL   Nature. 1997 Sep 18. 389(6648):300-5.
  PUBMED    9305847
REFERENCE   2  (bases 1 to 10024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            1997 Sep 18. 389(6648):300-5."
COMMENT     Sequence Label: pCEP4 Pig4
FEATURES             Location/Qualifiers
     source          1..10024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..132
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    275..409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(312..331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     366..385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(417..443)
                     /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="YAYDVPDYA"
     primer_bind     complement(839..858)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(840..858)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(915..939)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(936..1139)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(965..985)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1140..1519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2173..2221
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2246..2265
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2263..3300)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     promoter        complement(3343..3488)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     rep_origin      complement(3919..4507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3999..4018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4678..5469,5470..5538))
                     /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     5301..5320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5539..5643)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5711..5729
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      7785..9574
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
ORIGIN
        1 tctagagtcg acctcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
       61 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      121 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagccgga tcataatcag
      181 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
      241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
      301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
      361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc ttagctagcg
      421 taatctggaa catcgtaagc gtaatctgga acatcgtacc ccccgtattt ctcagacagg
      481 ccagcaggtc ggaagtgatt ggggtctttg ccactcctgc cccattcatt ggcagcctga
      541 tcagccagcg agtcctccgc accatggcca aagaatctct ggatattctc tctggcatcg
      601 ctgatcactt ctgcagccca ggcaccccca ggtccccttt tggcagcatc atagttcccc
      661 cgagcatgga agtatttgtc tgagccgatg taattggctt ctctcatgtc agagtaggct
      721 ctccacatgt cccgagcccc atcaaaagcc tcgccaagga acgaaaagaa gcttcggctg
      781 ctgacaccca ggaccaagga gcagaaaacc aggcccgtga gaagctccat ggtggtctcc
      841 ctatagtgag tcgtattagg atccctcgag cggccgctag caagcttgct agcagctggt
      901 acccagcttc tagagatctg acggttcact aaacgagctc tgcttatata gacctcccac
      961 cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca ttttggaaag
     1021 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     1081 aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc
     1141 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc
     1201 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga
     1261 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     1321 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     1381 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     1441 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     1501 ctagtcaata atcaatgtca acatggcggt catattggac atgagccaat ataaatgtac
     1561 atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata
     1621 taaccaatga ctaatatggc taattgccaa tattgattca atgtatagat cttccatacc
     1681 taccagttct gcgcctgcag caatggcaac aacgttgccc ggatctgcga tgataagctg
     1741 tcaaacatga gaattggtcg actagcttgg cacgccagaa atccgcgcgg tggtttttgg
     1801 gggtcggggg tgtttggcag ccacagacgc ccggtgttcg tgtcgcgcca gtacatgcgg
     1861 tccatgccca ggccatccaa aaaccatggg tctgtctgct cagtccagtc gtggacctga
     1921 ccccacgcaa cgcccaaaat aataaccccc acgaaccata aaccattccc catgggggac
     1981 cccgtcccta acccacgggg ccagtggcta tggcagggcc tgccgccccg acgttggctg
     2041 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg
     2101 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa
     2161 ccccgcgttt atgaacaaac gacccaacac ccgtgcgttt tattctgtct ttttattgcc
     2221 gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccat
     2281 ctcccgatcc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca
     2341 gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc
     2401 tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc
     2461 gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg
     2521 cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca
     2581 agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat
     2641 cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga
     2701 gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag
     2761 ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg
     2821 atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat
     2881 tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc
     2941 atccatggcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg
     3001 caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc
     3061 aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg
     3121 atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac
     3181 atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct
     3241 gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat
     3301 atctcattgc cccccgggat ctgcggcacg ctgttgacgc tgttaagcgg gtcgctgcag
     3361 ggtcgctcgg tgttcgaggc cacacgcgtc accttaatat gcgaagtgga cctcggaccg
     3421 cgccgccccg actgcatctg cgtgttcgaa ttcgccaatg acaagacgct gggcggggtt
     3481 tgtgtcatca tagaactaaa gacatgcaaa tatatttctt ccggggacac cgccagcaaa
     3541 cgcgagcaac gggccacggg gatgaagcag ggcggcacct cgctaacgga ttcaccactc
     3601 caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc
     3661 agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcggggccga
     3721 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat
     3781 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca
     3841 ggtagatgac gaccatcagg gacagcttca aggccagcaa aaggccagga accgtaaaaa
     3901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4321 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4981 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5281 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5641 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5701 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct catgtttgac
     5761 agcttatcat cgataagctg atcctcacag gccgcaccca gcttttcttc cgttgcccca
     5821 gtagcatctc tgtctggtga ccttgaagag gaagaggagg ggtcccgaga atccccatcc
     5881 ctaccgtcca gcaaaaaggg ggacgaggaa tttgaggcct ggcttgaggc tcaggacgca
     5941 aatcttgagg atgttcagcg ggagttttcc gggctgcgag taattggtga tgaggacgag
     6001 gatggttcgg aggatgggga attttcagac ctggatctgt ctgacagcga ccatgaaggg
     6061 gatgagggtg ggggggctgt tggagggggc aggagtctgc actccctgta ttcactgagc
     6121 gtcgtctaat aaagatgtct attgatctct tttagtgtga atcatgtctg acgaggggcc
     6181 aggtacagga cctggaaatg gcctaggaga gaagggagac acatctggac cagaaggctc
     6241 cggcggcagt ggacctcaaa gaagaggggg tgataaccat ggacgaggac ggggaagagg
     6301 acgaggacga ggaggcggaa gaccaggagc cccgggcggc tcaggatcag ggccaagaca
     6361 tagagatggt gtccggagac cccaaaaacg tccaagttgc attggctgca aagggaccca
     6421 cggtggaaca ggagcaggag caggagcggg aggggcagga gcaggagggg caggagcagg
     6481 aggaggggca ggagcaggag gaggggcagg agcaggaggg gcaggagcag gaggtggagg
     6541 ccggggtcga ggaggcagtg gaggccgggg tcgaggaggt agtggaggcc ggggtcgagg
     6601 aggtagtgga ggccgccggg gtagaggacg tgaaagagcc agggggggaa gtcgtgaaag
     6661 agccaggggg agaggtcgtg gacgtggaga aaagaggccc aggagtccca gtagtcagtc
     6721 atcatcatcc gggtctccac cgcgcaggcc ccctccaggt agaaggccat ttttccaccc
     6781 tgtaggggaa gccgattatt ttgaatacca ccaagaaggt ggcccagatg gtgagcctga
     6841 cgtgcccccg ggagcgatag agcagggccc cgcagatgac ccaggagaag gcccaagcac
     6901 tggaccccgg ggtcagggtg atggaggcag gcgcaaaaaa ggagggtggt ttggaaagca
     6961 tcgtggtcaa ggaggttcca acccgaaatt tgagaacatt gcagaaggtt taagagctct
     7021 cctggctagg agtcacgtag aaaggactac cgacgaagga acttgggtcg ccggtgtgtt
     7081 cgtatatgga ggtagtaaga cctcccttta caacctaagg cgaggaactg cccttgctat
     7141 tccacaatgt cgtcttacac cattgagtcg tctccccttt ggaatggccc ctggacccgg
     7201 cccacaacct ggcccgctaa gggagtccat tgtctgttat ttcatggtct ttttacaaac
     7261 tcatatattt gctgaggttt tgaaggatgc gattaaggac cttgttatga caaagcccgc
     7321 tcctacctgc aatatcaggg tgactgtgtg cagctttgac gatggagtag atttgcctcc
     7381 ctggtttcca cctatggtgg aaggggctgc cgcggagggt gatgacggag atgacggaga
     7441 tgaaggaggt gatggagatg agggtgagga agggcaggag tgatgtaact tgttaggaga
     7501 cgccctcaat cgtattaaaa gccgtgtatt cccccgcact aaagaataaa tccccagtag
     7561 acatcatgcg tgctgttggt gtatttctgg ccatctgtct tgtcaccatt ttcgtcctcc
     7621 caacatgggg caattgggca tacccatgtt gtcacgtcac tcagctccgc gctcaacacc
     7681 ttctcgcgtt ggaaaacatt agcgacattt acctggtgag caatcagaca tgcgacggct
     7741 ttagcctggc ctccttaaat tcacctaaga atgggagcaa ccagcaggaa aaggacaagc
     7801 agcgaaaatt cacgccccct tgggaggtgg cggcatatgc aaaggatagc actcccactc
     7861 tactactggg tatcatatgc tgactgtata tgcatgagga tagcatatgc tacccggata
     7921 cagattagga tagcatatac tacccagata tagattagga tagcatatgc tacccagata
     7981 tagattagga tagcctatgc tacccagata taaattagga tagcatatac tacccagata
     8041 tagattagga tagcatatgc tacccagata tagattagga tagcctatgc tacccagata
     8101 tagattagga tagcatatgc tacccagata tagattagga tagcatatgc tatccagata
     8161 tttgggtagt atatgctacc cagatataaa ttaggatagc atatactacc ctaatctcta
     8221 ttaggatagc atatgctacc cggatacaga ttaggatagc atatactacc cagatataga
     8281 ttaggatagc atatgctacc cagatataga ttaggatagc ctatgctacc cagatataaa
     8341 ttaggatagc atatactacc cagatataga ttaggatagc atatgctacc cagatataga
     8401 ttaggatagc ctatgctacc cagatataga ttaggatagc atatgctatc cagatatttg
     8461 ggtagtatat gctacccatg gcaacattag cccaccgtgc tctcagcgac ctcgtgaata
     8521 tgaggaccaa caaccctgtg cttggcgctc aggcgcaagt gtgtgtaatt tgtcctccag
     8581 atcgcagcaa tcgcgcccct atcttggccc gcccacctac ttatgcaggt attccccggg
     8641 gtgccattag tggttttgtg ggcaagtggt ttgaccgcag tggttagcgg ggttacaatc
     8701 agccaagtta ttacaccctt attttacagt ccaaaaccgc agggcggcgt gtgggggctg
     8761 acgcgtgccc ccactccaca atttcaaaaa aaagagtggc cacttgtctt tgtttatggg
     8821 ccccattggc gtggagcccc gtttaatttt cgggggtgtt agagacaacc agtggagtcc
     8881 gctgctgtcg gcgtccactc tctttcccct tgttacaaat agagtgtaac aacatggttc
     8941 acctgtcttg gtccctgcct gggacacatc ttaataaccc cagtatcata ttgcactagg
     9001 attatgtgtt gcccatagcc ataaattcgt gtgagatgga catccagtct ttacggcttg
     9061 tccccacccc atggatttct attgttaaag atattcagaa tgtttcattc ctacactagt
     9121 atttattgcc caaggggttt gtgagggtta tattggtgtc atagcacaat gccaccactg
     9181 aaccccccgt ccaaatttta ttctgggggc gtcacctgaa accttgtttt cgagcacctc
     9241 acatacacct tactgttcac aactcagcag ttattctatt agctaaacga aggagaatga
     9301 agaagcaggc gaagattcag gagagttcac tgcccgctcc ttgatcttca gccactgccc
     9361 ttgtgactaa aatggttcac taccctcgtg gaatcctgac cccatgtaaa taaaaccgtg
     9421 acagctcatg gggtgggaga tatcgctgtt ccttaggacc cttttactaa ccctaattcg
     9481 atagcatatg cttcccgttg ggtaacatat gctattgaat tagggttagt ctggatagta
     9541 tatactacta cccgggaagc atatgctacc cgtttagggt taacaagggg gccttataaa
     9601 cactattgct aatgccctct tgagggtccg cttatcggta gctacacagg cccctctgat
     9661 tgacgttggt gtagcctccc gtagtcttcc tgggcccctg ggaggtacat gtcccccagc
     9721 attggtgtaa gagcttcagc caagagttac acataaaggc aatgttgtgt tgcagtccac
     9781 agactgcaaa gtctgctcca ggatgaaagc cactcagtgt tggcaaatgt gcacatccat
     9841 ttataaggat gtcaactaca gtcagagaac ccctttgtgt ttggtccccc cccgtgtcac
     9901 atgtggaaca gggcccagtt ggcaagttgt accaaccaac tgaagggatt acatgcactg
     9961 ccccgcgaag aaggggcaga gatgtcgtag tcaggtttag ttcgtccggg gcggggatcg
    10021 atcc
//
