LOCUS       40924_49382        8366 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8366)
  AUTHORS   el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, 
            Lin D, Mercer WE, Kinzler KW, Vogelstein B
  TITLE     WAF1, a potential mediator of p53 tumor suppression.
  JOURNAL   Cell. 1993 Nov 19. 75(4):817-25.
  PUBMED    8242752
REFERENCE   2  (bases 1 to 8366)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8366)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1993 
            Nov 19. 75(4):817-25."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    complement(291..425)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(850..870)
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     intron          complement(1000..1065)
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(1199..2848)
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     2902..2918
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2918..2934
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5306..5322)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5355..5373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5383..5399)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5541..5996
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6022..6126
                     /label=AmpR promoter
     promoter        6446..6548
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6549..7205
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      7597..8185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8339..8356
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gatccagaca
      301 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      361 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      421 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      481 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat ctctagtcaa
      541 ggcactatac atcaaatatt ccttattaac ccctttacaa attaaaaagc taaaggtaca
      601 caatttttga gcatagttat taatagcaga cactctatgc ctgtgtggag taagaaaaaa
      661 cagtatgtta tgattataac tgttatgcct acttataaag gttacagaat atttttccat
      721 aattttcttg tatagcagtg cagctttttc ctttgtggtg taaatagcaa agcaagcaag
      781 agttctatta ctaaacacag catgactcaa aaaacttagc aattctgaag gaaagtcctt
      841 ggggtcttct acctttctct tcttttttgg aggagtagaa tgttgagagt cagcagtagc
      901 ctcatcatca ctagatggca tttcttctga gcaaaacagg ttttcctcat taaaggcatt
      961 ccaccactgc tcccattcat cagttccata ggttggaatc taaaatacac aaacaattag
     1021 aatcagtagt ttaacacatt atacacttaa aaattttata tttaccttag agctttaaat
     1081 ctctgtaggt agtttgtcca attatgtcac accacagaag taaggttcct tcacaaagat
     1141 cctctagagt attacaatag ctaagaattt cgtcatcgct gaatacagtt acattttaca
     1201 atttggactt tccgcccttc ttggccttta tgaggatctc tctgattttt cttgcgtcga
     1261 gttttccggt aagacctttc ggtacttcgt ccacaaacac aactcctccg cgcaactttt
     1321 tcgcggttgt tacttgactg gcgacgtaat ccacgatctc tttttccgtc atcgtctttc
     1381 cgtgctccaa aacaacaacg gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg
     1441 ccacgcccgc gtcgaagatg ttggggtgtt gtaacaatat cgattccaat tcagcggggg
     1501 ccacctgata tcctttgtat ttaattaaag acttcaagcg gtcaactatg aagaagtgtt
     1561 cgtcttcgtc ccagtaagct atgtctccag aatgtagcca tccatccttg tcaatcaagg
     1621 cgttggtcgc ttccggattg tttacataac cggacataat cataggtcct ctgacacata
     1681 attcgcctct ctgattaacg cccagcgttt tcccggtatc cagatccaca accttcgctt
     1741 caaaaaatgg aacaacttta ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca
     1801 gaatagctga tgtagtctca gtgagcccat atccttgtcg tatccctgga agatggaagc
     1861 gttttgcaac cgcttccccg acttctttcg aaagaggtgc gcccccagaa gcaatttcgt
     1921 gtaaattaga taaatcgtat ttgtcaatca gagtgctttt ggcgaagaat gaaaataggg
     1981 ttggtactag caacgcactt tgaattttgt aatcctgaag ggatcgtaaa aacagctctt
     2041 cttcaaatct atacattaag acgactcgaa atccacatat caaatatccg agtgtagtaa
     2101 acattccaaa accgtgatgg aatggaacaa cacttaaaat cgcagtatcc ggaatgattt
     2161 gattgccaaa aataggatct ctggcatgcg agaatctgac gcaggcagtt ctatgcggaa
     2221 gggccacacc cttaggtaac ccagtagatc cagaggaatt cattatcagt gcaattgttt
     2281 tgtcacgatc aaaggactct ggtacaaaat cgtattcatt aaaaccggga ggtagatgag
     2341 atgtgacgaa cgtgtacatc gactgaaatc cctggtaatc cgttttagaa tccatgataa
     2401 taattttctg gattattggt aatttttttt gcacgttcaa aattttttgc aacccctttt
     2461 tggaaacaaa cactacggta ggctgcgaaa tgttcatact gttgagcaat tcacgttcat
     2521 tataaatgtc gttcgcgggc gcaactgcaa ctccgataaa taacgcgccc aacaccggca
     2581 taaagaattg aagagagttt tcactgcata cgacgattct gtgatttgta ttcagcccat
     2641 atcgtttcat agcttctgcc aaccgaacgg acatttcgaa gtattccgcg tacgtgatgt
     2701 tcacctcgat atgtgcatct gtaaaagcaa ttgttccagg aaccagggcg tatctcttca
     2761 tagccttatg cagttgctct ccagcggttc catcctctag aggatagaat ggcgccgggc
     2821 ctttctttat gtttttggcg tcttccattt taccaacagt accggaatgc caagctcgag
     2881 ctccaccgcg gtggcggccg ctctagaact agtggatcga ggtcgacggt atcgataagc
     2941 ttctgacttc ggcagctgct cacacctcag ctggcgcagc tctgcgcggc cctgatatac
     3001 aaccgccccg cccgggcccg cctcaaggag gcgggacccg cgctcggccc accgcgccgc
     3061 ccgggaccgc cctccccggg gcccaggctc ctggctgccc agcgcggagc cagctgagcc
     3121 tggccgagtt ccagcaggcc agccggtccc ggaacctcgc gtgctgcaga ggggcacttc
     3181 cctcctcccc cagtccctcg cctgcgttgg tgcgctggac acatttcccc acgaagtgag
     3241 ccacaaatct ggcttttttt acttggagaa tgagttggca ctctccagga ggacacagca
     3301 ctgttagaat gagccccctt tctggctcac cgctgaccca ctctggcagg caaggattta
     3361 cccaatgcag ctgaaaagat caggaggatg acattaatac ataaaaattc ataaattata
     3421 aaaacgatgc acctctctgc aatttccaga aaagccccac aatatcacct ctattcccac
     3481 tgatccctca ctaggtcacc tctcccagaa gcacctggag cacctagaca ccccaacaaa
     3541 gcatcttgag gccggaatga tttttcagtc ccgtttattt cacagatgag gaaattgagg
     3601 tccactgaac ttaagtatac aaagttgttg attgtcacat gcttccggga aggagggaat
     3661 tggagagact accaaaaaag ggcaacctga tctccaggga aacagaagaa ttggacattg
     3721 aaccaatctc cctacaccct acactcacct gaacagaaga aacccctgtg gttgcagcag
     3781 ctttgttggc caggaagggg aggatttgac gagtgagttg tctgtctcct gaatactccc
     3841 cacatagccc gtatacactg ctggggaaac tggggctcag agaagtctgg tgacctaact
     3901 cagatcatgc agcaaagaaa tgactatagt tggaacacag gacttttgcc tcctgcccgg
     3961 ggctctctgc ttgtcatcct ttatttctgt ggttccaaaa tgacaaaaat gccaaataac
     4021 cctcatttgc agatggttta tggagatgac gtaaataaag gacaattctg gaagtgtcta
     4081 ctggttcttc tgacagacac agaaatgagt gatgtgccta tccgctccca tctacctcac
     4141 acccctgacc tcccctggac ttcacctttg cctcctttct gtgcctgaaa catttgcagt
     4201 tttgctttta aaaaattgca gaggatggat tgttcatctg aacagaaatc ccactgaaaa
     4261 acagaaccca ggcttggagc agctacaatt actgacatct caggctgctc agagtctgga
     4321 aatctctgcc cagacatgcc cagtcttctt cctctaacgc agctgacctc ggggaggaca
     4381 ggcttctttc tcctctgctg tggggatggg agcccaggta ggggtgggag gacacagcag
     4441 caggccccct tggcctgcct cgccctggag tcaggccagg attgtggcta aaccccagaa
     4501 gggccacagc acagcctcag gaccccactc taagccccac tgctgtctgc accttcgctc
     4561 ctatacatac aagccatcca aagggctggt tgtcaaatgt ccagcagagg acaggcacct
     4621 tctcccactg ccttgaagcc cctctgcttt caggcatttc aaatagacta gacattgtct
     4681 gtctgtttac ttcggggaga aatttttttt taatgaaggc ttgtaatgaa ttatttaaat
     4741 ttttgtggta ctgagcttca caatgaaata attctttttg ttgttgtttt tgggaaagaa
     4801 ttttttttat aacttctagc tcaccaccac cacgacattc aatatttctt aagtacctac
     4861 taagtgtcta gtactattca gtgctttaaa aaaaaaagtt tgcaaccatg cacttgaatg
     4921 tgtacccaga cacactctaa gggaggactt ctgccctgag gaaggcaagg gagcaggctg
     4981 ttaaaagtca cacacagaat ctgactccca gcacacactc acacaagcac acatgcatca
     5041 gatcccagcc ctgtcgcaag gatctgctgg cagatcacat accctgttca gagtaacagg
     5101 ctaaggctta cctggggcct ttagaggtct tctgtctcct accatcccct tcctcacctg
     5161 ataacaggca gcccaaggac aaaatagcca ccagcctctt ctatgccaga gctcaacatg
     5221 ttgggacatg ttcctaatgg ccagaaagcc aatcagagcc acagcctgct gcccaagctt
     5281 gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg
     5341 gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     5401 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5461 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5521 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5881 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5941 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     6001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6301 cgccccgaag aacgttttcc aatgatgagc acttttcgac cgaataaata cctgtgacgg
     6361 aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg aagccctggg
     6421 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     6481 atgaaataag atcactaccg ggcgtatttt ttgagttgtc gagattttca ggagctaagg
     6541 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     6601 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     6661 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     6721 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
     6781 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     6841 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     6901 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     6961 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     7021 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     7081 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     7141 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     7201 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     7261 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc cggtcgtcgg
     7321 ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac
     7381 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
     7441 ccccgtggag gtaataattg acgatatgat cctttttttc tgatcaaaaa ggatctaggt
     7501 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7561 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7621 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7681 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7741 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7801 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7861 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7921 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7981 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8041 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8101 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8161 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8221 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     8281 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     8341 cgagtcagtg agcgaggaag cggaag
//
