LOCUS       sequence-10519-e        6453 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 6453)
  AUTHORS   Irwin M, Marin MC, Phillips AC, Seelan RS, Smith DI, Liu W, Flores 
            ER, Tsai KY, Jacks T, Vousden KH, Kaelin WG
  TITLE     Role for the p53 homologue p73 in E2F-1-induced apoptosis.
  JOURNAL   Nature. 2000 Oct 5. 407(6804):645-8.
  PUBMED    11034215
REFERENCE   2  (bases 1 to 6453)
  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. 
            2000 Oct 5. 407(6804):645-8."
COMMENT     Sequence Label: T7-p73DD-pcDNA3
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2005..2023)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2006..2023)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2043..2060)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2049..2273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2319..2747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2406..2425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2616..2637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2756..2776)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2761..3091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2942..3077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3004..3023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3158..3952
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3212..3231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3822..3841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4126..4247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4163..4182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4217..4236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4296..4312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4296..4312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4309..4331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4320..4336
                     /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)."
     promoter        complement(join(4344..4350,4351..4368,4369..4374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4389..4410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4527..4544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4698..5286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4778..4797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5457..6248,6249..6317))
                     /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     6080..6099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6318..6422)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggctagca tgactggtgg acagcaaatg ggatccatga agcgtgcctt caagcagagc
      961 ccccctgccg tccccgccct tggtgccggt gtgaagaagc ggcggcatgg agacgaggac
     1021 acgtactacc ttcaggtgcg aggccgggag aactttgaga tcctgatgaa gctgaaagag
     1081 agcctggagc tgatggagtt ggtgccgcag ccactggtgg actcctatcg gcagcagcag
     1141 cagctcctac agaggccgag tcacctacag cccccgtcct acgggccggt cctctcgccc
     1201 atgaacaagg tgcacggggg catgaacaag ctgccctccg tcaaccagct ggtgggccag
     1261 cctcccccgc acagttcggc agctacaccc aacctggggc ccgtgggccc cgggatgctc
     1321 aacaaccatg gccacgcagt gccagccaac ggcgagatga gcagcagcca cagcgcccag
     1381 tccatggtct cggggtccca ctgcactccg ccacccccct accacgccga ccccagcctc
     1441 gtcagttttt taacaggatt ggggtgtcca aactgcatcg agtatttcac ctcccaaggg
     1501 ttacagagca tttaccacct gcagaacctg accattgagg acctgggggc cctgaagatc
     1561 cccgagcagt accgcatgac catctggcgg ggcctgcagg acctgaagca gggccacgac
     1621 tacagcaccg cgcagcagct gctccgctct agcaacgcgg ccaccatctc catcggcggc
     1681 tcaggggaac tgcagcgcca gcgggtcatg gaggccgtgc acttccgcgt gcgccacacc
     1741 atcaccatcc ccaaccgcgg cggcccaggc ggcggccctg acgagtgggc ggacttcggc
     1801 ttcgacctgc ccgactgcaa ggcccgcaag cagcccatca aggaggagtt cacggaggcc
     1861 gagatccact gagaattccc aattgcacct gaaattgcac tggaactgct gatggctgcg
     1921 aacttcttag attgttaagg tctccaattc tgcagatatc catcacactg gcggccgctc
     1981 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     2041 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2101 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2161 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2221 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2281 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2341 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2401 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2461 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2521 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2581 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2641 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     2701 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2761 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag
     2821 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2881 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2941 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3001 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3061 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3121 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3181 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3241 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3301 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3361 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3421 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3481 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3541 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3601 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3661 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3721 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3781 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3841 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3901 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3961 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4021 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4081 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4261 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4861 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     4921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5101 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5581 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6421 cgcacatttc cccgaaaagt gccacctgac gtc
//
