LOCUS       sequence-12186-e        5802 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 5802)
  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 5802)
  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-p53DD-pcDNA3
FEATURES             Location/Qualifiers
     source          1..5802
                     /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(1354..1372)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1355..1372)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1392..1409)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1398..1622
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1668..2096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1755..1774)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1965..1986
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2105..2125)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2110..2440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2291..2426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2353..2372
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2507..3301
                     /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(2561..2580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3171..3190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3475..3596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3512..3531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3566..3585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3645..3661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3645..3661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3658..3680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3669..3685
                     /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(3693..3699,3700..3717,3718..3723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3738..3759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3876..3893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4047..4635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4127..4146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4806..5597,5598..5666))
                     /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     5429..5448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5667..5771)
                     /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 ggatcccccc cagggagcac taagcgagca
      961 ctgcccaaca acaccagctc ctctccccag ccaaagaaga aaccactgga tggagaatat
     1021 ttcacccttc agatccgtgg gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc
     1081 ttggaactca aggatgccca ggctgggaag gagccagggg ggagcagggc tcactccagc
     1141 cacctgaagt ccaaaaaggg tcagtctacc tcccgccata aaaaacacat gttcaagaca
     1201 gaagggcctg actcagactg agaattccca attgcacctg aaattgcact ggaactgctg
     1261 atggctgcga acttcttaga ttgttaaggt ctccaattct gcagatatcc atcacactgg
     1321 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     1381 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1441 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1501 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1561 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1621 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1681 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1741 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1801 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     1861 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1921 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1981 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     2041 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2101 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     2161 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2281 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2341 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2401 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2461 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2521 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2581 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2641 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2701 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2761 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2821 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2881 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2941 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3001 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3061 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3121 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3181 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3241 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3301 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3361 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3421 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3601 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     4261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
