LOCUS       sequence-178011-        5672 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Luciferase reporter containing CDK4 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5672)
  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 5672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 active (880)
FEATURES             Location/Qualifiers
     source          1..5672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      138..147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             144..1796
                     /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(210..228)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1837..1958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1848..1867)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1902..1921
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2206..2223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2377..2965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2457..2476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3136..3927,3928..3996))
                     /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     3759..3778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3997..4101)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4128..4583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4215..4234)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4425..4446
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4714..4762
                     /note="synthetic polyadenylation signal"
     misc_feature    4776..4867
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4816..4835
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cacctcctgt ccgcccctca gcgcatgggt ggcggtcacg tgcccagaac gtccggcgtt
       61 cgccccgccc tcccagtttc cgcgcgcctc tttggcagct ggtcacataa gcttggcatt
      121 ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa ggcccggcgc
      181 cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat
      241 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt
      301 acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga
      361 atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt
      421 tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg
      481 aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt
      541 tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg
      601 attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac
      661 ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg
      721 cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata
      781 gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc
      841 cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac
      901 tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt
      961 ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct
     1021 ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg
     1081 cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc
     1141 tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac
     1201 ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg
     1261 ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga
     1321 gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg
     1381 acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct
     1441 tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg
     1501 aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc
     1561 ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga
     1621 tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc
     1681 gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa
     1741 gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaattct
     1801 agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat gagtttggac
     1861 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1921 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1981 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     2041 aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt caacccagtc
     2101 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     2161 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
     2221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     4141 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4201 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4261 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4321 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4381 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4441 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4501 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4561 caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact gttgggaagg
     4621 gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat aagtaagtaa
     4681 tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt ttcattacat
     4741 ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa
     4801 acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca
     4861 tttctctatc gataggtacc gagctcttac gcgtgctagc ccgggctcga gaaggtcaag
     4921 tattcctgtc tcctgctcat caataacttc tctgccagat ggcctcaggt ctttgcatta
     4981 acactcccag ggaggtttgg aagttggaaa agtgagagga ttaccttgtc actgagcctg
     5041 ttggattact gtattaatct gccttccgtc atcctttaat cgtagttttg ccctcaaatg
     5101 ggcattgttt actccctatt gtagaagacc ccattgttgg ctacatcttt atagatagtc
     5161 gtaccactac tttaaagggc cagtgacaga caccaaaaac aaaataaaaa cccacataca
     5221 cactggaagc aagcactcag taaaatgttg taggattaat gaatgaaaca atgacacaaa
     5281 tgagagtatg ttttctcatg gtggagcgaa aaggtgacag catcacctct ggtaccccaa
     5341 ctcccacccc ctccccaatg cagacaggct gaaagaccgg tagtgagact ggagttcagc
     5401 cttcagaccg gtagtgagac aatccttcag ccgggagttg ggctctgggt ggcctaggtt
     5461 gccatggcac cgcctcgggc tccaccctct cttgtccccc tcaccgctcc ccccctgcag
     5521 cgggggttgt ggcagccagt cacgtgcccg ccgcgtagcc acacctctgc tcctcagagc
     5581 aatgtcaagc ggtcacgtgt gatagcaaca gatcacgtgg ctgccatcgc ccctccgccc
     5641 ccttacactc ttcgccctcc tcccagtcga ag
//
