LOCUS       sequence-365348-        5894 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 5894)
  AUTHORS   Boyd SD, Tsai KY, Jacks T
  TITLE     An intact HDM2 RING-finger domain is required for nuclear exclusion 
            of p53.
  JOURNAL   Nat Cell Biol. 2000 Sep . 2(9):563-8.
  PUBMED    10980695
REFERENCE   2  (bases 1 to 5894)
  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. 2000 Sep . 2(9):563-8."
COMMENT     Sequence Label: GFP-p53 (NES-)
FEATURES             Location/Qualifiers
     source          1..5894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             43..837
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             43..837
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(97..116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     707..726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1025..1044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1069..1116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1445..2033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1934..1953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2208..2511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2512..2715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2666..2686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2787..3965
                     /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
                     FRELNEAAEAKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     regulatory      3981..3990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3987..3989,3990..3992,3993..4706)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3987..3989,3990..3992,3993..4703)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4032..4053)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4293..4312)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4640..4661
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4829..4950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4866..4885)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4920..4939
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4957..5412)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5094..5115)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5306..5325
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5439..5543
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5545..5894,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5545..5736
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5568..5588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5753..5888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
       61 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
      121 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
      181 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
      241 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
      301 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
      361 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
      421 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
      481 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
      541 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
      601 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
      661 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
      721 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
      781 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
      841 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
      901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
      961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     1021 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1081 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     1141 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     1201 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     1261 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     1321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1561 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2101 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     2161 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2221 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2281 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2341 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2401 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2461 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2521 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     2581 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     2641 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     2701 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
     2761 tcgagctcaa gcttcgaatt ctgcagatgg aggagccgca gtcagatcct agcgtcgagc
     2821 cccctctgag tcaggaaaca ttttcagacc tatggaaact acttcctgaa aacaacgttc
     2881 tgtccccctt gccgtcccaa gcaatggatg atttgatgct gtccccggac gatattgaac
     2941 aatggttcac tgaagaccca ggtccagatg aagctcccag aatgccagag gctgctcccc
     3001 gcgtggcccc tgcaccagca gctcctacac cggcggcccc tgcaccagcc ccctcctggc
     3061 ccctgtcatc ttctgtccct tcccagaaaa cctaccaggg cagctacggt ttccgtctgg
     3121 gcttcttgca ttctgggaca gccaagtctg tgacttgcac gtactcccct gccctcaaca
     3181 agatgttttg ccaactggcc aagacctgcc ctgtgcagct gtgggttgat tccacacccc
     3241 cgcccggcac ccgcgtccgc gccatggcca tctacaagca gtcacagcac atgacggagg
     3301 ttgtgaggcg ctgcccccac catgagcgct gctcagatag cgatggtctg gcccctcctc
     3361 agcatcttat ccgagtggaa ggaaatttgc gtgtggagta tttggatgac agaaacactt
     3421 ttcgacatag tgtggtggtg ccctatgagc cgcctgaggt tggctctgac tgtaccacca
     3481 tccactacaa ctacatgtgt aacagttcct gcatgggcgg catgaaccgg aggcccatcc
     3541 tcaccatcat cacactggaa gactccagtg gtaatctact gggacggaac agctttgagg
     3601 tgcgtgtttg tgcctgtcct gggagagacc ggcgcacaga ggaagagaat ctccgcaaga
     3661 aaggggagcc tcaccacgag ctgcccccag ggagcactaa gcgagcactg cccaacaaca
     3721 ccagctcctc tccccagcca aagaagaaac cactggatgg agaatatttc acccttcaga
     3781 tccgtgggcg tgagcgcttc gagatgttcc gagagctgaa tgaggccgcg gaagccaagg
     3841 atgcccaggc tgggaaggag ccagggggga gcagggctca ctccagccac ctgaagtcca
     3901 aaaagggtca gtctacctcc cgccataaaa aactcatgtt caagacagaa gggcctgact
     3961 cagaccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     4021 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     4081 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4141 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     4201 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4261 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4321 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4381 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4441 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     4501 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4561 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     4621 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4681 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     4741 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4801 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4861 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4921 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4981 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5041 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5101 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5161 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5221 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5281 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5341 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5401 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5461 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5521 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     5581 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5641 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5701 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5761 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5821 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5881 tggaggccta ggct
//
