LOCUS Exported 5039 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Luciferase reporter containing CDK4 promoter. ACCESSION . VERSION . KEYWORDS CDk4-luciferase fragment 4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5039) TITLE Modiano Lab plasmids REFERENCE 2 (bases 1 to 5039) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5039 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..1653 /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(67..85) /label=LucNrev /note="Firefly luciferase, reverse primer" polyA_signal 1694..1815 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1705..1724) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1759..1778 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2063..2080) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2234..2822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2314..2333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2993..3853) /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 3616..3635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3854..3958) /gene="bla" /label=AmpR promoter rep_origin 3985..4440 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4072..4091) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4282..4303 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 4571..4619 /note="synthetic polyadenylation signal" misc_feature 4633..4724 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 4673..4692 /label=RVprimer3 /note="pGL3 vector, forward primer" regulatory 5034..4 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga 61 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt 121 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc 181 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta 241 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt 301 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt 361 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa 421 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga 481 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat 541 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga 601 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg 661 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt 721 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt 781 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac 841 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg 901 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct 961 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat 1021 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc 1081 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa 1141 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt 1201 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct 1261 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct 1321 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa 1381 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt 1441 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat 1501 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac 1561 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata 1621 aaggccaaga agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt 1681 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga 1741 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc 1801 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag 1861 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat 1921 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg 1981 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt 2041 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 2101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 2281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 2341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 2401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 2461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 2521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 2581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 2641 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 2701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 2761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 2821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 2881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 2941 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 3001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 3061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 3121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 3181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 3241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 3301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 3361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 3421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 3481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 3541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 3601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 3661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 3721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 3781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 3841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 3901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 3961 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg 4021 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 4081 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 4141 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 4201 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 4261 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 4321 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa 4381 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt 4441 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc 4501 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg 4561 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga 4621 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa 4681 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag 4741 ctcttacgcg tgctagcccg ggctcgagaa ggtcaagtat tcctgtctcc tgctcatcaa 4801 taacttctct gccagatggc ctcaggtctt tgcattaaca ctcccaggga ggtttggaag 4861 ttggaaaagt gagaggatta ccttgtcact gagcctgttg gattactgta ttaatctgcc 4921 ttccgtcatc ctttaatcgt agttttgccc tcaaatgggc attgtttact ccctattgta 4981 gaagacccca ttgttggcta catcaagctt ggcattccgg tactgttggt aaagccacc //