LOCUS       Exported                7824 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Neo-Bam p53 R273H
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7824)
  AUTHORS   Baker SJ, Markowitz S, Fearon ER, Willson JK, Vogelstein B
  TITLE     Suppression of human colorectal carcinoma cell growth by wild-type 
            p53.
  JOURNAL   Science. 1990 Aug 24. 249(4971):912-5.
  PUBMED    2144057
REFERENCE   2  (bases 1 to 7824)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7824
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        468..671
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     668..692
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          783..1355
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1485..2666
                     /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
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVHVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALSNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(2833..2852)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2898..2953
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2952..2971)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3311..3327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3311..3327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3324..3346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3335..3351
                     /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(3359..3389)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3404..3425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3542..3559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3713..4301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3793..3812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4472..5332)
                     /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     5095..5114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5333..5437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5505..5523
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5561..5583)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5683..5702
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5896..5918
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5910..5927
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5911..5927
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6338..6483
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6540..7334
                     /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(6594..6613)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7204..7223
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7528..7547)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7572..7619
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ctagagagct tggcccattg catacgttgt atccatatca taatatgtac atttatattg
       61 gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat taatagtaat
      121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
      721 acctccatag aagacaccgg gaccgatcca gcctccggtc gaccgatcct gagaacttca
      781 gggtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat
      841 atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc
      901 atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc
      961 ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac
     1021 gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt
     1081 tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag agaacaattg
     1141 ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata
     1201 ttcttattgg tagaaacaac tacaccctgg tcatcatcct gcctttctct ttatggttac
     1261 aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg
     1321 ctaaccatgt tcatgccttc ttctctttcc tacagctcct gggcaacgtg ctggttgttg
     1381 tgctgtctca tcattttggc aaagaattcc tcgacggatc cgaattccac gacggtgaca
     1441 cgcttccctg gattggcagc cagactgcct tccgggtcac tgccatggag gagccgcagt
     1501 cagatcctag cgtcgagccc cctctgagtc aggaaacatt ttcagaccta tggaaactac
     1561 ttcctgaaaa caacgttctg tcccccttgc cgtcccaagc aatggatgat ttgatgctgt
     1621 ccccggacga tattgaacaa tggttcactg aagacccagg tccagatgaa gctcccagaa
     1681 tgccagaggc tgctccccgc gtggcccctg caccagcagc tcctacaccg gcggcccctg
     1741 caccagcccc ctcctggccc ctgtcatctt ctgtcccttc ccagaaaacc taccagggca
     1801 gctacggttt ccgtctgggc ttcttgcatt ctgggacagc caagtctgtg acttgcacgt
     1861 actcccctgc cctcaacaag atgttttgcc aactggccaa gacctgccct gtgcagctgt
     1921 gggttgattc cacacccccg cccggcaccc gcgtccgcgc catggccatc tacaagcagt
     1981 cacagcacat gacggaggtt gtgaggcgct gcccccacca tgagcgctgc tcagatagcg
     2041 atggtctggc ccctcctcag catcttatcc gagtggaagg aaatttgcgt gtggagtatt
     2101 tggatgacag aaacactttt cgacatagtg tggtggtgcc ctatgagccg cctgaggttg
     2161 gctctgactg taccaccatc cactacaact acatgtgtaa cagttcctgc atgggcggca
     2221 tgaaccggag gcccatcctc accatcatca cactggaaga ctccagtggt aatctactgg
     2281 gacggaacag ctttgaggtg catgtttgtg cctgtcctgg gagagaccgg cgcacagagg
     2341 aagagaatct ccgcaagaaa ggggagcctc accacgagct gcccccaggg agcactaagc
     2401 gagcactgtc caacaacacc agctcctctc cccagccaaa gaagaaacca ctggatggag
     2461 aatatttcac ccttcagatc cgtgggcgtg agcgcttcga gatgttccga gagctgaatg
     2521 aggccttgga actcaaggat gcccaggctg ggaaggagcc aggggggagc agggctcact
     2581 ccagccacct gaagtccaaa aagggtcagt ctacctcccg ccataaaaaa ctcatgttca
     2641 agacagaagg gcctgactca gactgacatt ctccacttct tgttccccac tgacagcctc
     2701 ccacccccat ctctccctcc cctgccattt tgaattcgga tccgtcgagg aattcactcc
     2761 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     2821 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     2881 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2941 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     3001 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     3061 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     3121 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     3181 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     3241 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacggat cggccgcaat
     3301 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3361 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4141 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5461 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5521 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5581 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5641 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     5701 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5761 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     5821 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     5881 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttcacg
     5941 ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag
     6001 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     6061 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     6121 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     6181 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     6241 caggggatca agatcctgct tcatccccgt ggcccgttgc tcgcgtttgc tggcggtgtc
     6301 cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc cgcccagcgt
     6361 cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc ccaggtccac
     6421 ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca gcgacccgct
     6481 taacagcgtc aacagcgtgc cgcagatctg atcaagagac aggatgagga tcgtttcgca
     6541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6721 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     7081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7321 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     7381 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7441 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     7501 ccaccccggg agatggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     7561 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     7621 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     7681 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     7741 cccagggctc gcagccaacg tcggggcggc aagccctgcc atagccacgg gccccgtggg
     7801 ttagggacgg cggatcgcgg ccct
//
