LOCUS       sequence-215676-        8111 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 8111)
  AUTHORS   Yu J, Zhang L, Hwang PM, Rago C, Kinzler KW, Vogelstein B
  TITLE     Identification and classification of p53-regulated genes.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Dec 7. 96(25):14517-22.
  PUBMED    10588737
REFERENCE   2  (bases 1 to 8111)
  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: "Proc Natl 
            Acad Sci U S A. 1999 Dec 7. 96(25):14517-22."
COMMENT     Sequence Label: pBI-p73 wt/EGFP
FEATURES             Location/Qualifiers
     source          1..8111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(112..130,131..153,154..172,173..195,196..214,215..237,
                     238..256,257..279,280..298,299..321,322..340,341..363,
                     364..382)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        415..453
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             768..794
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          3596..4168
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4304..4323)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4369..4424
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4423..4442)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4822..4839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4993..5581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5073..5092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5752..6543,6544..6612))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6375..6394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6613..6717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6785..6803
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    7166..7300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7203..7222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7257..7276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7337..8050,8051..8053,8054..8056))
                     /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             complement(join(7340..8050,8051..8053,8054..8056))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7382..7403)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     7731..7750
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7990..8011
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8064..8080)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gctggatcgg tcccggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc
       61 gatctgacgg ttcactaaac gagctctgct tatataggtc gagtttacca ctccctatca
      121 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
      181 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
      241 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      301 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      361 cactccctat cagtgataga gaaaagtgaa agtcgagctc ggtacccggg tcgaggtagg
      421 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      481 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
      541 ggccccgaat tcgagctcgg tacccgggga tcctctagtc agagcttgtc gacggtaccc
      601 cggggaattc ggcagatccc gaggtccgac agcgcccggc ccagatcccc acgcctgcca
      661 ggagcaagcc gagagccagc cggccggcgc actccgactc cgagcagtct ctgtccttcg
      721 acccgagccc cgcgcccttt ccgggacccc tgccccgcgg atccatgtac ccttacgatg
      781 taccggatta cgcactcgag gcccagtcca ccaccacctc ccccgatggg ggcaccacgt
      841 ttgagcacct ctggagctct ctggaaccag acagcaccta cttcgacctt ccccagtcaa
      901 gccgggggaa taatgaggtg gtgggtggca cggattccag catggacgtc ttccacctag
      961 agggcatgac cacatctgtc atggcccagt tcaatttgct gagcagcacc atggaccaga
     1021 tgagcagccg cgctgcctcg gccagcccgt acaccccgga gcacgccgcc agcgtgccca
     1081 cccattcacc ctacgcacag cccagctcca ccttcgacac catgtcgccc gcgcctgtca
     1141 tcccctccaa caccgactat cccggacccc accacttcga ggtcactttc cagcagtcca
     1201 gcacggccaa gtcagccacc tggacgtact ccccactctt gaagaaactc tactgccaga
     1261 tcgccaagac atgccccatc cagatcaagg tgtccgcccc accgcccccg ggcaccgcca
     1321 tccgggccat gcctgtctac aagaaggcgg agcacgtgac cgacatcgtg aagcgctgcc
     1381 ccaaccacga gctcgggagg gacttcaacg aaggacagtc tgccccagcc agccacctca
     1441 tccgtgtgga aggcaataat ctctcgcagt atgtggacga ccctgtcacc ggcaggcaga
     1501 gcgtcgtggt gccctatgag ccaccacagg tggggacaga attcaccacc atcctgtaca
     1561 acttcatgtg taacagcagc tgtgtggggg gcatgaaccg acggcccatc ctcatcatca
     1621 tcaccctgga gacgcgggat gggcaggtgc tgggccgccg gtccttcgag ggccgcatct
     1681 gcgcctgtcc tggccgcgac cgaaaagccg atgaggacca ctaccgggag cagcaggcct
     1741 tgaatgagag ctccgccaag aacggggctg ccagcaagcg cgccttcaag cagagtcccc
     1801 ctgccgtccc cgccctgggc ccgggtgtga agaagcggcg gcacggagac gaggacacgt
     1861 actacctgca ggtgcgaggc cgcgagaact tcgagatcct gatgaagctg aaggagagcc
     1921 tggagctgat ggagttggtg ccgcagccgc tggtagactc ctatcggcag cagcagcagc
     1981 tcctacagag gccgagtcac ctacagcccc catcctacgg gccggtcctc tcgcccatga
     2041 acaaggtgca cgggggcgtg aacaagctgc cctccgtcaa ccagctggtg ggccagcctc
     2101 ccccgcacag ctcggcagct acacccaacc tgggacctgt gggctctggg atgctcaaca
     2161 accacggcca cgcagtgcca gccaacagcg agatgaccag cagccacggc acccagtcca
     2221 tggtctcggg gtcccactgc actccgccac ccccctacca cgccgacccc agcctcgtca
     2281 gttttttaac aggattgggg tgtccaaact gcatcgagta tttcacgtcc caggggttac
     2341 agagcattta ccacctgcag aacctgacca tcgaggacct gggggccctg aagatccccg
     2401 agcagtatcg catgaccatc tggcggggcc tgcaggacct gaagcagggc cacgactacg
     2461 gcgccgccgc gcagcagctg ctccgctcca gcaacgcggc cgccatttcc atcggcggct
     2521 ccggggagct gcagcgccag cgggtcatgg aggccgtgca cttccgcgtg cgccacacca
     2581 tcaccatccc caaccgcggc ggccccggcg ccggccccga cgagtgggcg gacttcggct
     2641 tcgacctgcc cgactgcaag gcccgcaagc agcccatcaa ggaggagttc acggaggccg
     2701 agatccactg aggggccggg cccagccaga gcctgtgcca ccgcccagag acccaggccg
     2761 cctcgctctc cttcctgtgt ccaaaactgc ctccggaggc agggcctcca ggctgtgccc
     2821 ggggaaaggc aaggtccggc ccatgccccg gcacctcacc ggccccagga gaggcccagc
     2881 caccaaagcc gcctgcggac agcctgagtc acctgcagaa ccttctggag ctgccctaat
     2941 gctgggcttg cggggcaggg gccggcccac tctcagccct gccactgccg ggcgtgctcc
     3001 atggcaggcg tgggtgggga ccgcagtgtc agctccgacc tccaggcctc atcctagaga
     3061 ctctgtcatc tgccgatcaa gcaaggtcct tccagaggaa agaatcctct tcgctggtgg
     3121 actgccaaaa agtattttgc gacatctttt ggttctggag agtggtgagc agccaagcga
     3181 ctgtgtctga aacaccgtgc attttcaggg aatgtcccta acgggctggg gactctctct
     3241 gctggacttg ggagtggcct ttgcccccag cacactgtat tctgcgggac cgcctccttc
     3301 ctgcccctaa caaccaccaa agtgttgctg aaattggaga aaactgggga aggcgcaacc
     3361 cctcccaggt gcgggaagca tctggtaccg cctcggccag tgcccctcag cctggccaca
     3421 gtcacctctc cttggggaac cctgggcaga aagggacagc ctgtccttag aggaccggaa
     3481 attgtcaata tttgataaaa tgataccctt ttctacaaaa aaaaaaaaaa aaagggcccg
     3541 gatccactag ttctagcgcg gcctcgacga tatctctaga gctgagaact tcagggtgag
     3601 tttggggacc cttgattgtt ctttcttttt cgctattgta aaattcatgt tatatggagg
     3661 gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc accatggacc
     3721 ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt ctcctcttat
     3781 tttcttttca ttttctgtaa ctttttcgtt aaactttagc ttgcatttgt aacgaatttt
     3841 taaattcact tttgtttatt tgtcagattg taagtacttt ctctaatcac ttttttttca
     3901 aggcaatcag ggtatattat attgtacttc agcacagttt tagagaacaa ttgttataat
     3961 taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa atattcttat
     4021 tggtagaaac aactacaccc tggtcatcat cctgcctttc tctttatggt tacaatgata
     4081 tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct ctgctaacca
     4141 tgttcatgcc ttcttctctt tcctacagct cctgggcaac gtgctggttg ttgtgctgtc
     4201 tcatcatttt ggcaaagaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
     4261 tggtgtggcc aatgccctgg ctcacaaata ccactgagat cgatcttttt ccctctgcca
     4321 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     4381 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     4441 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     4501 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     4561 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     4621 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     4681 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     4741 tatggagatc cctcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4801 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4861 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4921 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4981 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5041 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5101 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5161 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5221 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5281 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5341 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5401 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5461 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5521 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5581 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5641 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5701 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5761 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5821 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5881 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5941 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6001 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6061 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6121 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6181 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6241 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6301 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6361 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6421 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6481 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6541 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6601 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6661 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6721 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6781 taaaaatagg cgtatcacga ggccctttcg tcttcactcg acgcggccag cttggtcgag
     6841 ctggatactt cccgtccgcc agggggacat gccggcgatg ctgaaggtcg cgcgcattcc
     6901 cgatgaagag gccggttacc gcctgttgac ctggtgggac gggcagggcg ccgcccgagt
     6961 cttcgcctcg gcggcgggcg ctctgctcat ggagcgcgcg tccggggccg gggaccttgc
     7021 acagatagcg tggtccggcc aggacgacga ggcttgcagg atcataatca gccataccac
     7081 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     7141 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     7201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7261 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc ctctagaact aggtcgacaa
     7321 gcttgatatc gaattcttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     7381 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     7441 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     7501 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     7561 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     7621 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     7681 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     7741 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     7801 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     7861 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     7921 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     7981 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     8041 cgcccttgct caccatccgc ggggatccac tagttctaga gcggccgctg caggaattcg
     8101 gggccgcgga g
//
