LOCUS Exported 7401 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION luciferase assay. ACCESSION . VERSION . KEYWORDS hsa-cdk2 3'UTR sicheck2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7401) AUTHORS Hsu AY, Wang D, Liu S, Lu J, Syahirah R, Bennin DA, Huttenlocher A, Umulis DM, Wan J, Deng Q TITLE Phenotypical microRNA screen reveals a noncanonical role of CDK2 in regulating neutrophil migration. JOURNAL Proc Natl Acad Sci U S A. 2019 Aug 26. pii: 1905221116. doi: 10.1073/pnas.1905221116. PUBMED 31451657 REFERENCE 2 (bases 1 to 7401) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7401 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 5..362 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(28..48) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 213..348 /label=SV40 ori /note="SV40 origin of replication" primer_bind 275..294 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 432..564 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 547..566 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 609..627 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 631..640 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 637..1572 /codon_start=1 /product="Renilla luciferase" /label=hRluc /note="human codon-optimized" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" polyA_signal 2759..2807 /note="synthetic polyadenylation signal" promoter 2816..3567 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" regulatory 3597..3606 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3603..5255 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /db_xref="GI:42718124" /protein_id="AAS38485.1" /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" RBS 4562..4570 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" polyA_signal 5299..5420 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5310..5329) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5364..5383 /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter 5553..5657 /gene="bla" /label=AmpR promoter CDS 5658..6518 /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(5876..5895) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6689..7277 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7178..7197 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc 61 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 181 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 241 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 301 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 361 aaaagcttga ttcttctgac acaacagtct cgaacttaag ctgcagaagt tggtcgtgag 421 gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg 481 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca 541 tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc tcttaaggct 601 agagtactta atacgactca ctataggcta gccaccatgg cttccaaggt gtacgacccc 661 gagcaacgca aacgcatgat cactgggcct cagtggtggg ctcgctgcaa gcaaatgaac 721 gtgctggact ccttcatcaa ctactatgat tccgagaagc acgccgagaa cgccgtgatt 781 tttctgcatg gtaacgctgc ctccagctac ctgtggaggc acgtcgtgcc tcacatcgag 841 cccgtggcta gatgcatcat ccctgatctg atcggaatgg gtaagtccgg caagagcggg 901 aatggctcat atcgcctcct ggatcactac aagtacctca ccgcttggtt cgagctgctg 961 aaccttccaa agaaaatcat ctttgtgggc cacgactggg gggcttgtct ggcctttcac 1021 tactcctacg agcaccaaga caagatcaag gccatcgtcc atgctgagag tgtcgtggac 1081 gtgatcgagt cctgggacga gtggcctgac atcgaggagg atatcgccct gatcaagagc 1141 gaagagggcg agaaaatggt gcttgagaat aacttcttcg tcgagaccat gctcccaagc 1201 aagatcatgc ggaaactgga gcctgaggag ttcgctgcct acctggagcc attcaaggag 1261 aagggcgagg ttagacggcc taccctctcc tggcctcgcg agatccctct cgttaaggga 1321 ggcaagcccg acgtcgtcca gattgtccgc aactacaacg cctaccttcg ggccagcgac 1381 gatctgccta agatgttcat cgagtccgac cctgggttct tttccaacgc tattgtcgag 1441 ggagctaaga agttccctaa caccgagttc gtgaaggtga agggcctcca cttcagccag 1501 gaggacgctc cagatgaaat gggtaagtac atcaagagct tcgtggagcg cgtgctgaag 1561 aacgagcagt aattctaggc gatcgctcga gagccttctt gaagccccca gccctaatct 1621 caccctctcc tccagtgtgg gcttgaccag gcttggcctt gggctatttg gactcaggtg 1681 ggccctctga acttgcctta aacactcacc ttctagtctt ggccagccaa ctctgggaat 1741 acaggggtga aaggggggaa ccagtgaaaa tgaaaggaag tttcagtatt agatgcactt 1801 aagttagcct ccaccaccct ttcccccttc tcttagttat tgctgaagag ggttggtata 1861 aaaataattt taaaaaagcc ttcctacacg ttagatttgc cgtaccaatc tctgaatgcc 1921 ccataattat tatttccagt gtttgggatg accaggatcc caagcctcct gctgccacaa 1981 tgtttataaa ggccaaatga tagcgggggc taagttggtg cttttgagaa ccaagtaaaa 2041 caaaaccact gggaggagtc tattttaaag aattcggttg aaaaaataga tccaatcagt 2101 ttatacccta gttagtgttt tgcctcacct aataggctgg gagactgaag actcagcccg 2161 ggtggggctg cagaaaaatg attggcccca gtccccttgt ttgtcccttc tacaggcatg 2221 aggaatctgg gaggccctga gacagggatt gtgcttcatt ccaatctatt gcttcaccat 2281 ggccttatga ggcaggtgag agatgtttga atttttctct tccttttagt attcttagtt 2341 gttcagttgc caaggatccc tgatcccatt ttcctctgac gtccacctcc taccccatag 2401 gagttagaag ttagggttta ggcatcattt tgagaatgct gacacttttt cagggctgtg 2461 attgagtgag ggcatgggta aaaatatttc tttaaaagaa ggatgaacaa ttatatttat 2521 atttcaggtt atatccaata gtagagttgg cttttttttt ttttggtcat agtgggtgga 2581 tttgttgcca tgtgcacctt ggggttttgt aatgacagtg ctaaaaaaaa aagcattttt 2641 ttttatgatt tgtctctgtc acccttgtcc ttgagtgctc ttgctattaa cgttatttgt 2701 aatttagttt gtagctcatt aaaaaaatgt gcctagtttt atagcggccg ctggccgcaa 2761 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgagg atctaaatga 2821 gtcttcggac ctcgcggggg ccgcttaagc ggtggttagg gtttgtctga cgcgggggga 2881 gggggaagga acgaaacact ctcattcgga ggcggctcgg ggtttggtct tggtggccac 2941 gggcacgcag aagagcgccg cgatcctctt aagcaccccc ccgccctccg tggaggcggg 3001 ggtttggtcg gcgggtggta actggcgggc cgctgactcg ggcgggtcgc gcgccccaga 3061 gtgtgacctt ttcggtctgc tcgcagaccc ccgggcggcg ccgccgcggc ggcgacgggc 3121 tcgctgggtc ctaggctcca tggggaccgt atacgtggac aggctctgga gcatccgcac 3181 gactgcggtg atattaccgg agaccttctg cgggacgagc cgggtcacgc ggctgacgcg 3241 gagcgtccgt tgggcgacaa acaccaggac ggggcacagg tacactatct tgtcacccgg 3301 aggcgcgagg gactgcagga gcttcaggga gtggcgcagc tgcttcatcc ccgtggcccg 3361 ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat ttgcatgtct ttagttctat 3421 gatgacacaa accccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc 3481 ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac 3541 cgagcgaccc tgcagcgacc cgcttaaaag cttggcattc cggtactgtt ggtaaagcca 3601 ccatggccga tgctaagaac attaagaagg gccctgctcc cttctaccct ctggaggatg 3661 gcaccgctgg cgagcagctg cacaaggcca tgaagaggta tgccctggtg cctggcacca 3721 ttgccttcac cgatgcccac attgaggtgg acatcaccta tgccgagtac ttcgagatgt 3781 ctgtgcgcct ggccgaggcc atgaagaggt acggcctgaa caccaaccac cgcatcgtgg 3841 tgtgctctga gaactctctg cagttcttca tgccagtgct gggcgccctg ttcatcggag 3901 tggccgtggc ccctgctaac gacatttaca acgagcgcga gctgctgaac agcatgggca 3961 tttctcagcc taccgtggtg ttcgtgtcta agaagggcct gcagaagatc ctgaacgtgc 4021 agaagaagct gcctatcatc cagaagatca tcatcatgga ctctaagacc gactaccagg 4081 gcttccagag catgtacaca ttcgtgacat ctcatctgcc tcctggcttc aacgagtacg 4141 acttcgtgcc agagtctttc gacagggaca aaaccattgc cctgatcatg aacagctctg 4201 ggtctaccgg cctgcctaag ggcgtggccc tgcctcatcg caccgcctgt gtgcgcttct 4261 ctcacgcccg cgaccctatt ttcggcaacc agatcatccc cgacaccgct attctgagcg 4321 tggtgccatt ccaccacggc ttcggcatgt tcaccaccct gggctacctg atttgcggct 4381 ttcgggtggt gctgatgtac cgcttcgagg aggagctgtt cctgcgcagc ctgcaagact 4441 acaaaattca gtctgccctg ctggtgccaa ccctgttcag cttcttcgct aagagcaccc 4501 tgatcgacaa gtacgacctg tctaacctgc acgagattgc ctctggcggc gccccactgt 4561 ctaaggaggt gggcgaagcc gtggccaagc gctttcatct gccaggcatc cgccagggct 4621 acggcctgac cgagacaacc agcgccattc tgattacccc agagggcgac gacaagcctg 4681 gcgccgtggg caaggtggtg ccattcttcg aggccaaggt ggtggacctg gacaccggca 4741 agaccctggg agtgaaccag cgcggcgagc tgtgtgtgcg cggccctatg attatgtccg 4801 gctacgtgaa taaccctgag gccacaaacg ccctgatcga caaggacggc tggctgcact 4861 ctggcgacat tgcctactgg gacgaggacg agcacttctt catcgtggac cgcctgaagt 4921 ctctgatcaa gtacaagggc taccaggtgg ccccagccga gctggagtct atcctgctgc 4981 agcaccctaa cattttcgac gccggagtgg ccggcctgcc cgacgacgat gccggcgagc 5041 tgcctgccgc cgtcgtcgtg ctggaacacg gcaagaccat gaccgagaag gagatcgtgg 5101 actatgtggc cagccaggtg acaaccgcca agaagctgcg cggcggagtg gtgttcgtgg 5161 acgaggtgcc caagggcctg accggcaagc tggacgcccg caagatccgc gagatcctga 5221 tcaaggctaa gaaaggcggc aagatcgccg tgtaataatt ctagagtcgg ggcggccggc 5281 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 5341 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 5401 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 5461 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata 5521 aggatccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 5581 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 5641 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 5701 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 5761 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 5821 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 5881 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 5941 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 6001 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 6061 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 6121 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 6181 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 6241 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 6301 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 6361 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 6421 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 6481 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 6541 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 6601 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 6661 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 6721 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 6781 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 6841 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 6901 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 6961 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 7021 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 7081 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 7141 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 7201 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 7261 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 7321 ggccttttgc tcacatggct cgacagatct gcgcagcacc atggcctgaa ataacctctg 7381 aaagaggaac ttggttaggt a //