LOCUS       sequence-473443-        7160 bp    DNA     circular SYN 21-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7160)
  AUTHORS   el-Deiry WS, Tokino T, Waldman T, Oliner JD, Velculescu VE, Burrell 
            M, Hill DE, Healy E, Rees JL, Hamilton SR
  TITLE     Topological control of p21WAF1/CIP1 expression in normal and 
            neoplastic tissues.
  JOURNAL   Cancer Res. 1995 Jul 1. 55(13):2910-9.
  PUBMED    7796420
REFERENCE   2  (bases 1 to 7160)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer Res.
            1995 Jul 1. 55(13):2910-9."
COMMENT     Sequence Label: p21 promoter A
FEATURES             Location/Qualifiers
     source          1..7160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1044..1061)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1044..1060)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1053..1075)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1269..1288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1388..1410
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1448..1466)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1534..1638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1639..1707,1708..2499)
                     /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     complement(1857..1876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2670..3258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3159..3178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3412..3429
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3546..3567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3582..3587,3588..3605,3606..3612)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3620..3636
                     /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)."
     primer_bind     3625..3647
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3644..3660
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3644..3660
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(3688..4201)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3883..3904)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4095..4114
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4216..4350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4240..4259)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4294..4313
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4775..4795)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4925..4990
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5157..5816)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5731..5750
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aacagaaatc ccactgaaaa acagaaccca ggcttggagc agctacaatt actgacatct
       61 caggctgctc agagtctgga aatctctgcc cagacatgcc cagtcttctt cctctaacgc
      121 agctgacctc ggggaggaca ggcttctttc tcctctgctg tggggatggg agcccaggta
      181 ggggtgggag gacacagcag cagaccccct tggcctgcct cgccctggag tcaggccagg
      241 attgtggcta aaccccagaa gggccacagc acagcctcag gaccccactc taagccccac
      301 tgctgtctgc accttcgctc ctatacatac aaaccatcca aagggctggt tgtcaaatgt
      361 ccagcagagg acaggcacct tctcccactg ccttgaagcc cctctgcttt caggcatttc
      421 aaatagacta gacattgtct gtctgtttac tttggggaga aaattttttt taataaaggc
      481 ttgtaatgaa ttatttaaat ttttgtggta ctgagcttca caatgaaata attctttttg
      541 ttgttgtttt tgggaaagaa ttttttttat aacttctagc tcaccaccac cacgacattc
      601 aatatttctt aagtacctac taagtgtcta gtactattca gtgctttaaa aaaaaaagtt
      661 tgcaaccatg cacttgaatg tgtacccaga cacactctaa gggaggactt ctgccctgaa
      721 gaaggcaagg gagcaggctg ttaaaagtca cacacagaat ctgactccca gcacacactc
      781 acacaagcac acatgcatta gatcccagcc ctgtcgcaag gatctgctgg cagatcacat
      841 accctgttca gagtaacagg ctaaggttta cctggggtct ttagaggtct cctgtctcct
      901 accatcccct tcctcacctg aaaacaggca gcccaaggac aaaatagcca ccagcctctt
      961 ctatgccaga gctcaacatg ttgggacatg ttcctgatgg ccagaaagcc aatcagagcc
     1021 acagcctgct gcccaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1081 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1141 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1201 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1261 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1321 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1381 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     1441 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1501 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1861 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2341 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3241 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3301 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3361 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3421 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3481 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3541 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3601 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3661 atgattacga attcgagctc ggtacccacc ccggttgata atcagaaaag ccccaaaaac
     3721 aggaagattg tataagcaaa tatttaaatt gtaagcgtta atattttgtt aaaattcgcg
     3781 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3841 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3901 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3961 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4021 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4081 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4141 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4201 tggggatcgg ggatcgatcc agacatgata agatacattg atgagtttgg acaaaccaca
     4261 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4321 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4381 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4441 atggctgatt atgatctcta gtcaaggcac tatacatcaa atattcctta ttaacccctt
     4501 tacaaattaa aaagctaaag gtacacaatt tttgagcata gttattaata gcagacactc
     4561 tatgcctgtg tggagtaaga aaaaacagta tgttatgatt ataactgtta tgcctactta
     4621 taaaggttac agaatatttt tccataattt tcttgtatag cagtgcagct ttttcctttg
     4681 tggtgtaaat agcaaagcaa gcaagagttc tattactaaa cacagcatga ctcaaaaaac
     4741 ttagcaattc tgaaggaaag tccttggggt cttctacctt tctcttcttt tttggaggag
     4801 tagaatgttg agagtcagca gtagcctcat catcactaga tggcatttct tctgagcaaa
     4861 acaggttttc ctcattaaag gcattccacc actgctccca ttcatcagtt ccataggttg
     4921 gaatctaaaa tacacaaaca attagaatca gtagtttaac acattataca cttaaaaatt
     4981 ttatatttac cttagagctt taaatctctg taggtagttt gtccaattat gtcacaccac
     5041 agaagtaagg ttccttcaca aagatccggc gaatttctgc cattcatccg cttattatca
     5101 cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac
     5161 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     5221 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     5281 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     5341 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     5401 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     5461 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     5521 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     5581 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     5641 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     5701 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     5761 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     5821 gcttccttag ctcctgaaaa tctcgccaag ctcctcgagg aactgacttc ggcagctgct
     5881 cacacctcag ctggcgcagc tcagcgcggc cctgatatac aaccgccccg cccgggcccg
     5941 cctcaaggag gcgggacccg cgctcggccc accgcgccgc ccgggaccgc cctccccggg
     6001 gcccaggctc ctggctgccc agcgcggagc cagctgagcc tggccgagtt ccagcaggcc
     6061 agccggtccc ggaacctcgc gtgctgcagg agggcacttc cctcctcccc cagtccctcg
     6121 cctgcgttgg tgcgctggac acatttcccc acgaagtgag ccacaaatct ggcttttttt
     6181 acttggagaa tgagttggca ctctccagga ggacacagca ctgttagaat gagccccctt
     6241 tctggctcac cgctgaccca ctctggcagg caaggattta cccaatgcag ctgaaaagat
     6301 caggaggatg acattaatac ataaaaattc ataaattata aaaacgatgc acctctctgc
     6361 aatttccaga aaagccccac aatatcacct ctattcccac tgatccctca ctaggtcacc
     6421 tctcccagaa gcacctggag cacctagaca ccccaacaaa gcatcttgag gccagaatga
     6481 tttttcagtc ccgtttattt cacagatgag gaaattgagg tccactgaac ttaagtatac
     6541 aaagttgttg attgtcacat gcttccggga aggagggaat tggagagact accaaaaaag
     6601 ggcaacctga tctccaggga aacagaagaa ttggacattg aaccaatctc cctacgccct
     6661 acactcacct gaacagaaga aacccctgtg gttgcagcag ctttgttggc caggaagggg
     6721 aggatttgac gagtgagttg tctgtctcct gaatactccc cacatagccc gtatacactg
     6781 ctggggaaac tggggctcag agaagtctgg tgacctaact cagatcatgc agcaaagaaa
     6841 tgactatagt tggaacacag gacttttgcc tcctgcccgg ggctctctgc ttgtcatcct
     6901 ttatttctgt ggttccaaaa tgacaaaaat gtcaaataac cctcatttgc agatggttta
     6961 tggagatgac ataaataaag gacgattctg gaagtgtcta ctggttcttc tgacagacac
     7021 agaaatgagt gatgtgtcta tccgctccca tctacctcac acccctgacc tcccctggac
     7081 ttcacctttg cctcctttct gtgcctgaaa catttgcagt tttgctttta aaaaattgca
     7141 gaggatggat tgttcatctg
//
