LOCUS       Exported                7900 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Neo-Bam p53 R175H
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7900)
  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 7900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7900
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1410..2591
                     /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
                     KQSQHMTEVVRHCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(2770..2789)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2835..2890
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2889..2908)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3248..3264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3248..3264)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3261..3283)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3272..3288
                     /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(3296..3326)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3341..3362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3531..3548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3702..4290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3782..3801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4461..5321)
                     /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     5084..5103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5322..5426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5494..5512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5550..5572)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5672..5691
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5885..5907
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5899..5916
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5900..5916
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6327..6472
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6529..7323
                     /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(6583..6602)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7193..7212
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7517..7536)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7561..7608
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atcccccggg ctgcaggaat tccacgacgg tgacacgctt ccctggattg
     1381 gcagccagac tgccttccgg gtcactgcca tggaggagcc gcagtcagat cctagcgtcg
     1441 agccccctct gagtcaggaa acattttcag acctatggaa actacttcct gaaaacaacg
     1501 ttctgtcccc cttgccgtcc caagcaatgg atgatttgat gctgtccccg gacgatattg
     1561 aacaatggtt cactgaagac ccaggtccag atgaagctcc cagaatgcca gaggctgctc
     1621 cccgcgtggc ccctgcacca gcagctccta caccggcggc ccctgcacca gccccctcct
     1681 ggcccctgtc atcttctgtc ccttcccaga aaacctacca gggcagctac ggtttccgtc
     1741 tgggcttctt gcattctggg acagccaagt ctgtgacttg cacgtactcc cctgccctca
     1801 acaagatgtt ttgccaactg gccaagacct gccctgtgca gctgtgggtt gattccacac
     1861 ccccgcccgg cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag cacatgacgg
     1921 aggttgtgag gcactgcccc caccatgagc gctgctcaga tagcgatggt ctggcccctc
     1981 ctcagcatct tatccgagtg gaaggaaatt tgcgtgtgga gtatttggat gacagaaaca
     2041 cttttcgaca tagtgtggtg gtgccctatg agccgcctga ggttggctct gactgtacca
     2101 ccatccacta caactacatg tgtaacagtt cctgcatggg cggcatgaac cggaggccca
     2161 tcctcaccat catcacactg gaagactcca gtggtaatct actgggacgg aacagctttg
     2221 aggtgcgtgt ttgtgcctgt cctgggagag accggcgcac agaggaagag aatctccgca
     2281 agaaagggga gcctcaccac gagctgcccc cagggagcac taagcgagca ctgcccaaca
     2341 acaccagctc ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc
     2401 agatccgtgg gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca
     2461 aggatgccca ggctgggaag gagccggggg ggagcagggc tcactccagc cacctgaagt
     2521 ccaaaaaggg tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg
     2581 actcagactg acattctcca cttcttgttc cccactgaca gcctcccacc cccatctctc
     2641 cctcccctgc cattttgaat tcctgcagcc cgggggatcc gtcgaggaat tcactcctca
     2701 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     2761 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     2821 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2881 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     2941 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     3001 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     3061 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     3121 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     3181 cccagtcata gctgtccctc ttctcttatg gagatccctc gacggatcgg ccgcaattcg
     3241 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3301 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3361 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3421 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctca ttaatgaatc
     3481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4141 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5461 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5521 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5581 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5641 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5701 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     5761 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5821 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     5881 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttcacgc tgccgcaagc
     5941 actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac
     6001 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     6061 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     6121 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     6181 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     6241 gatcctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa
     6301 tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg
     6361 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta
     6421 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aacagcgtca
     6481 acagcgtgcc gcagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     6541 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6601 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6661 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6721 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6781 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6841 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6901 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6961 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     7021 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     7081 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     7141 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     7201 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     7261 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     7321 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     7381 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     7441 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccggga
     7501 gatgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     7561 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     7621 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     7681 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     7741 cagccaacgt cggggcggca agccctgcca tagccacggg ccccgtgggt tagggacggc
     7801 ggatcgcggc cctctagaga gcttggccca ttgcatacgt tgtatccata tcataatatg
     7861 tacatttata ttggctcatg tccaacatta ccgccatgtt
//
