LOCUS       sequence-140269-        5025 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 5025)
  AUTHORS   Hermeking H, Rago C, Schuhmacher M, Li Q, Barrett JF, Obaya AJ, 
            O'Connell BC, Mateyak MK, Tam W, Kohlhuber F, Dang CV, Sedivy JM, 
            Eick D, Vogelstein B, Kinzler KW
  TITLE     Identification of CDK4 as a target of c-MYC.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 Feb 29. 97(5):2229-34.
  PUBMED    10688915
REFERENCE   2  (bases 1 to 5025)
  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: "Proc Natl 
            Acad Sci U S A. 2000 Feb 29. 97(5):2229-34."
COMMENT     Sequence Label: pBV-Luc wt MBS1-4
FEATURES             Location/Qualifiers
     source          1..5025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      289..298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             295..1947
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(361..379)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1988..2109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1999..2018)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2053..2072
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2357..2374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2528..3116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2608..2627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3287..4078,4079..4147))
                     /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     3910..3929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4148..4252)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4279..4734
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4366..4385)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4576..4597
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4865..4913
                     /note="synthetic polyadenylation signal"
     misc_feature    4927..5018
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4967..4986
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgggt tgttggcagc cagtcacgtg cccgccgcgt agccacacct ctgctcctca
       61 gagcaatgtc aagcggtcac gtgtgatagc aacagatcac gtggctgcca tcgcccctcc
      121 gcccccttac actcttcgcc ctcctcccag tcgaagcacc tcctgtccgc ccctcagcgc
      181 catgggtggc ggtcacgtgc ccagaacgtc cggaattcgg gtatataatg gatccggtat
      241 cgagatctgc gatctaagta agcttggcat tccggtactg ttggtaaagc caccatggaa
      301 gacgccaaaa acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct
      361 ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt
      421 acagatgcac atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg
      481 ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt
      541 gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt
      601 gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag
      661 cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag
      721 ctcccaatca tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag
      781 tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg
      841 ccagagtcct tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact
      901 ggtctgccta aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc
      961 agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca
     1021 ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc
     1081 gtcttaatgt atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt
     1141 caaagtgcgc tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac
     1201 aaatacgatt tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa
     1261 gtcggggaag cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc
     1321 actgagacta catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc
     1381 ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg
     1441 ggcgttaatc aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta
     1501 aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac
     1561 atagcttact gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt
     1621 aagtacaaag gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc
     1681 aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc
     1741 gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc
     1801 gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta
     1861 ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc
     1921 aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg gcggccggcc gcttcgagca
     1981 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2041 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2101 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2161 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg
     2221 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2281 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2341 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4261 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4321 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4381 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4441 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4501 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4561 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4621 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4681 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt
     4741 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4801 gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg
     4861 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4921 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc
     4981 tgtccccagt gcaagtgcag gtgccagaac atttctctat cgata
//
