LOCUS       sequence-272722-        5871 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 5871)
  AUTHORS   Jiang P, Du W, Wang X, Mancuso A, Gao X, Wu M, Yang X
  TITLE     p53 regulates biosynthesis through direct inactivation of 
            glucose-6-phosphate dehydrogenase.
  JOURNAL   Nat Cell Biol. 2011 Mar;13(3):310-6. Epub 2011 Feb 20.
  PUBMED    21336310
REFERENCE   2  (bases 1 to 5871)
  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: "Nat Cell 
            Biol."; date: "2011-03"; volume: "13(3)"; pages: "310-6. Epub 2011 
            Feb 20"
COMMENT     Sequence Label: Flag-p53/pRK5
FEATURES             Location/Qualifiers
     source          1..5871
                     /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"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      912..921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             921..944
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             951..2129
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     regulatory      2158..2167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2171..2305
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2208..2227)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2262..2281
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2374..2731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2374..2565
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2397..2417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2582..2717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2751..2768)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2751..2767)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2760..2782)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2980..3435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3067..3086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3277..3298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3452..3471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3571..3593
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3631..3649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3717..3821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3822..3890,3891..4682)
                     /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     complement(4040..4059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4853..5441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5342..5361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5595..5612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5729..5750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5765..5770,5771..5788,5789..5795)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5803..5819
                     /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     5808..5830
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5827..5843
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5827..5843
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
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 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cgccgccatg gactacaagg acgacgatga caagggatcc atggaggagc
      961 cgcagtcaga tcctagcgtc gagccccctc tgagtcagga aacattttca gacctatgga
     1021 aactacttcc tgaaaacaac gttctgtccc ccttgccgtc ccaagcaatg gatgatttga
     1081 tgctgtcccc ggacgatatt gaacaatggt tcactgaaga cccaggtcca gatgaagctc
     1141 ccagaatgcc agaggctgct ccccccgtgg cccctgcacc agcagctcct acaccggcgg
     1201 cccctgcacc agccccctcc tggcccctgt catcttctgt cccttcccag aaaacctacc
     1261 agggcagcta cggtttccgt ctgggcttct tgcattctgg gacagccaag tctgtgactt
     1321 gcacgtactc ccctgccctc aacaagatgt tttgccaact ggccaagacc tgccctgtgc
     1381 agctgtgggt tgattccaca cccccgcccg gcacccgcgt ccgcgccatg gccatctaca
     1441 agcagtcaca gcacatgacg gaggttgtga ggcgctgccc ccaccatgag cgctgctcag
     1501 atagcgatgg tctggcccct cctcagcatc ttatccgagt ggaaggaaat ttgcgtgtgg
     1561 agtatttgga tgacagaaac acttttcgac atagtgtggt ggtgccctat gagccgcctg
     1621 aggttggctc tgactgtacc accatccact acaactacat gtgtaacagt tcctgcatgg
     1681 gcggcatgaa ccggaggccc atcctcacca tcatcacact ggaagactcc agtggtaatc
     1741 tactgggacg gaacagcttt gaggtgcgtg tttgtgcctg tcctgggaga gaccggcgca
     1801 cagaggaaga gaatctccgc aagaaagggg agcctcacca cgagctgccc ccagggagca
     1861 ctaagcgagc actgcccaac aacaccagct cctctcccca gccaaagaag aaaccactgg
     1921 atggagaata tttcaccctt cagatccgtg ggcgtgagcg cttcgagatg ttccgagagc
     1981 tgaatgaggc cttggaactc aaggatgccc aggctgggaa ggagccaggg gggagcaggg
     2041 ctcactccag ccacctgaag tccaaaaagg gtcagtctac ctcccgccat aaaaaactca
     2101 tgttcaagac agaagggcct gactcagacg tcgactgact cgacctgcag aagcttggcc
     2161 gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2221 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2281 caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat
     2341 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     2401 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2461 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2641 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2701 aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc
     2761 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2821 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2881 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2941 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3001 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3061 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3121 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3181 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3241 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3301 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3361 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3421 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3481 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3541 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3601 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3661 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3721 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3781 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3841 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3901 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3961 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4021 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4081 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4141 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4201 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4261 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4321 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4381 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4441 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4501 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4561 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4621 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4681 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4741 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4801 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4861 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4921 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4981 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5041 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5101 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5161 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5221 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5281 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5341 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5401 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5461 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5521 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5581 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5641 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5701 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     5761 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     5821 ttcacacagg aaacagctat gacatgatta cgaattaatt cgagctcgcc c
//
