LOCUS       sequence-84063-e        7092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CFP tagged p53 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7092)
  AUTHORS   Lahav G, Rosenfeld N, Sigal A, Geva-Zatorsky N, Levine AJ, Elowitz 
            MB, Alon U
  TITLE     Dynamics of the p53-Mdm2 feedback loop in individual cells.
  JOURNAL   Nat Genet. 2004 Feb;36(2):147-50. Epub 2004 Jan 18.
  PUBMED    14730303
REFERENCE   2  (bases 1 to 7092)
  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 
            Genet."; date: "2004-02"; volume: "36(2)"; pages: "147-50. Epub 2004
            Jan 18"
COMMENT     Sequence Label: Mtp-p53-ECFP
FEATURES             Location/Qualifiers
     source          1..7092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /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(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2223..2239
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2223..2239
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      2491..2500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2497..3675
                     /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
                     SPDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     CDS             join(3685..3687,3688..3690,3691..4404)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3730..3751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3991..4010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4338..4359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(4976..6001)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(4976..6001)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6075..6404)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6089..6224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6389..6409
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(6448..6467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6449..6467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6474..6491)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6474..6490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6483..6505)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6632..7087
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6719..6738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6929..6950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctcgaaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     2221 gctctagaac tagtggatcc cgctgcactc cgcccgaaaa gtgcgctcgg ctctgccaag
     2281 gacgcggggc gcgtgactat gcgtgggctg gagcaaccgc ctgctgggtg caaacccttt
     2341 gcgcccggac tcgtccaacg actataaaga gggcaggctg tcctctaagc gtcaccacga
     2401 cttcaacgtc ctgagtacct tctcctcact tactccgtag ctccagcttc accagatctc
     2461 gagctcaagc ttcgaattct gcagtcgacc gccaccatgg aggagccgca gtcagatcct
     2521 agcgtcgagc cccctctgag tcaggaaaca ttttcagacc tatggaaact acttcctgaa
     2581 aacaacgttc tgtccccctt gccgtcccaa gcaatggatg atttgatgct gtccccggac
     2641 gatattgaac aatggttcac tgaagaccca ggtccagatg aagctcccag aatgccagag
     2701 gctgctcccc gcgtggcccc tgcaccagca gctcctacac cggcggcccc tgcaccagcc
     2761 ccctcctggc ccctgtcatc ttctgtccct tcccagaaaa cctaccaggg cagctacggt
     2821 ttccgtctgg gcttcttgca ttctgggaca gccaagtctg tgacttgcac gtactcccct
     2881 gccctcaaca agatgttttg ccaactggcc aagacctgcc ctgtgcagct gtgggttgat
     2941 tccacacccc cgcccggcac ccgcgtccgc gccatggcca tctacaagca gtcacagcac
     3001 atgacggagg ttgtgaggcg ctgcccccac catgagcgct gctcagatag cgatggtctg
     3061 gcccctcctc agcatcttat ccgagtggaa ggaaatttgc gtgtggagta tttggatgac
     3121 agaaacactt ttcgacatag tgtggtggtg ccctatgagc cgcctgaggt tggctctgac
     3181 tgtaccacca tccactacaa ctacatgtgt aacagttcct gcatgggcgg catgaaccgg
     3241 aggcccatcc tcaccatcat cacactggag gactccagtg gtaatctact gggacggaac
     3301 agctttgagg tgcgtgtttg tgcctgtcct gggagagacc ggcgcacaga ggaagagaat
     3361 ctccgcaaga aaggggagcc tcaccacgag ctgcccccag ggagcactaa gcgagcactg
     3421 cccaacaaca ccagctcctc tccccagcca aagaagaaac cactggatgg agaatatttc
     3481 acccttcaga tccgtgggcg tgagcgcttc gagatgttcc gagagctgaa tgaggccttg
     3541 gaactcaagg atgcccaggc tgggaaggag ccagggggga gcagggctca ctccagccac
     3601 ctgaagtcca aaaagggtca gtctacctcc cgccataaaa aactcatgtt caagacagaa
     3661 gggcctgact cagacgggga tccaatggtg agcaagggcg aggagctgtt caccggggtg
     3721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3841 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
     3901 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     4021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4081 gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat
     4141 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     4201 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4261 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4321 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4381 ggcatggacg agctgtacaa gtaatccgga ctcagatctc ggaatggacc ccaactgctc
     4441 ctgctccacc ggcggctcct gcacttgcac cagctcctgc gcctgcaaga actgcaagtg
     4501 cacctcctgc aagaagagct gctgctcctg ctgtcccgtg ggctgctcca aatgtgccca
     4561 gggctgtgtc tgcaaaggcg ccgcggacaa gtgcacgtgc tgtgcctgat gtgacgaaca
     4621 gcgctgccac cacgtgtaaa tagtatcgga ccaacccagc gtcttcctat acagttccac
     4681 cctgtttact aaacccccgt tttctaccga gtacgtgaat aataaaagcc tgtttgagtc
     4741 taactctggt tttcttggtg tggtttggca ataagaaact ggggtgactt gatagtctgg
     4801 ggatctggtt ttggaccccc tcgtgccttt acctcccgcc tctggccctc acagaggggt
     4861 aatgtctttg ggtaaagcca agctatccca taagcttatc gatgccttgc taggctcgcg
     4921 cgagttgctg gctgaggcgt tctcgaaatc agctcttgtt cggtcggcat ctactctatt
     4981 cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac
     5041 agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg
     5101 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc
     5161 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc
     5221 gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac
     5281 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca
     5341 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg
     5401 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca
     5461 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt
     5521 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga
     5581 ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg
     5641 catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt
     5701 gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctaaattccc
     5761 caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac
     5821 gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta
     5881 catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc
     5941 tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca
     6001 tatctcattg cccccggacg agagatctgc agggcttccc aaccttacca gagggcgccc
     6061 cagctggcaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     6121 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     6181 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     6241 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     6301 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     6361 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca caggtcgacc
     6421 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     6481 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6541 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6601 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     6661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6841 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6961 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     7081 acaatttagg tg
//
