LOCUS       sequence-181629-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter containing CDK4 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  AUTHORS   Baksh S, Widlund HR, Frazer-Abel AA, Du J, Fosmire S, Fisher DE, 
            DeCaprio JA, Modiano JF, Burakoff SJ
  TITLE     NFATc2-mediated repression of cyclin-dependent kinase 4 expression.
  JOURNAL   Mol Cell. 2002 Nov;10(5):1071-81.
  PUBMED    12453415
REFERENCE   2  (bases 1 to 6584)
  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: "Mol Cell.";
            date: "2002-11"; volume: "10(5)"; pages: "1071-81"
COMMENT     Sequence Label: CDk4-luciferase fragment A
FEATURES             Location/Qualifiers
     source          1..6584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1815..3467
                     /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(1881..1899)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3508..3629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3519..3538)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3573..3592
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3877..3894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4048..4636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4128..4147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4807..5598,5599..5667))
                     /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     5430..5449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5668..5772)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5799..6254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5886..5905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6096..6117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6385..6433
                     /note="synthetic polyadenylation signal"
     misc_feature    6447..6538
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6487..6506
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gtgaacaata cacacactta cccctttcct tttatggaga cttccctctg aggccaagtt
       61 ctacccccat cacatttttt tctttcttac acttacattc tccctctcct cccctttgca
      121 aagctgaggg aaaagggatt ccaggggagc tcattgggtc aaacatactg ggaaaaaatt
      181 taattagtga aagagtaatt aacagaagaa gggaggggac agagcctggg acagactgcc
      241 aaattaaggt tcctcaatag aaagtgatgg agagacagta atgttcccat gaagtattca
      301 aatgttttgg atggactaat gaatgaatca gaaaaagggg ctgggaccca agcagacaga
      361 gaggacgtct tctgttgcaa agacagtaag ctgtgaaggt cttatgtgga aggtgaggct
      421 gaagtgggga atgctggtgt gggaagggag tttgaaaaac tcacagacaa ttatcttagc
      481 cttccagaag gcttacaaag cacccacccc tagcaaggta gccaataaaa cattcacctc
      541 ttgagcccct tagtcagttc agtccttgag gtttttctta ctagtgttcc agctgccctt
      601 gccaggaggg gcccagggct acaagtatca ggatggaaag aggaggcatc tgtattaata
      661 tgtattctat tttgctctaa gtcatggttt atattcatct tctagtttga actcttatat
      721 tctcagccag atggttaagg aaaacagaag ttagagtgga aggagtggca aatagggtaa
      781 taaaatggct ttagatttta gacaaattct cacaaatgag acactaaccg ctttcctcct
      841 ttactcccat acccccaaag ggctgggaga taaagacctt ggagccaata aaacaaagta
      901 ggagaaatct acttgtgtct acctaggtgt ctgggcagag gccatttggg aacaagtgtg
      961 ttctggacag tgctaagtgc taagaaagcg gcactaggga aaaagaggga agcacaataa
     1021 ctccaaatga gtagtcacaa ggtttgcaga agggggcatg tcattttcat tcattgtggt
     1081 ggtctggagg gtgcaggggg tgtggtggta actttcttgc acatgtgcaa tcctgtatcc
     1141 gtcttatata tgatgtgata aataatatca tatacataca tatatacata gaagcatata
     1201 tgcacacata caaatatcac ataatcatag aaccttccta ctggaaaaaa aaaaaaaact
     1261 taaaaggtca agtattcctg tctcctgctc atcaataact tctctgccag atggcctcag
     1321 gtctttgcat taacactccc agggaggttt ggaagttgga aaagtgagag gattaccttg
     1381 tcaccgagcc tgttggatta ctgtattaat ctgccttccg tcatccttta atcgtagttt
     1441 tgccctcaaa tgggcattgt ttactcccta ttgtagaaga ccccattgtt ggctacatct
     1501 ttatagatag tcgtaccact actttaaagg gccagtgaca gacaccaaaa acaaaataaa
     1561 aacccacata cacactggaa gcaagcactc agtaaaatgt tgtagaatta atgaatgaaa
     1621 caatgacaca aatgagagta tgttttctca tggtggagcg aaaaggtgac agcatcacct
     1681 ctggtacccc aactcccacc ccctccccaa tgcagacagg ctgaaagacc ggtagtgaga
     1741 ctggagttca gcctcagacc ggtagtgaga caatccttca gccgggagtt gggctctggg
     1801 tggcctaggt tgccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
     1861 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
     1921 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     1981 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     2041 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     2101 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     2161 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     2221 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     2281 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     2341 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     2401 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     2461 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     2521 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     2581 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     2641 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     2701 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     2761 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     2821 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     2881 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     2941 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     3001 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     3061 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     3121 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     3181 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     3241 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     3301 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     3361 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     3421 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg
     3481 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3541 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3601 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3661 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3721 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     3781 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     3841 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     5821 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5881 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5941 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6001 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6061 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6121 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6181 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6241 aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6301 gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt
     6361 acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     6421 ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
     6481 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat
     6541 cgataggtac cgagctctta cgcgtgctag cccgggctcg agaa
//
