LOCUS       sequence-237981-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  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 7406)
  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. 2019 Aug 26. pii: 1905221116. doi: 
            10.1073/pnas.1905221116."
COMMENT     Sequence Label: hsa-cdk2 3'UTR sicheck2 mut
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        59..250
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(82..102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      267..402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          486..618
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     601..620
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        663..681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      685..694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             691..1626
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2818..2866
                     /note="synthetic polyadenylation signal"
     promoter        2875..3626
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3656..3665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3662..5314
                     /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             4621..4629
                     /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    5358..5479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5369..5388)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5423..5442
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5612..5716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5717..5785,5786..6577)
                     /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(5935..5954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6748..7336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7237..7256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
       61 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
      121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
      181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
      361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
      421 ttgattcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg tgaggcactg
      481 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
      541 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
      601 ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta
      661 cttaatacga ctcactatag gctagccacc atggcttcca aggtgtacga ccccgagcaa
      721 cgcaaacgca tgatcactgg gcctcagtgg tgggctcgct gcaagcaaat gaacgtgctg
      781 gactccttca tcaactacta tgattccgag aagcacgccg agaacgccgt gatttttctg
      841 catggtaacg ctgcctccag ctacctgtgg aggcacgtcg tgcctcacat cgagcccgtg
      901 gctagatgca tcatccctga tctgatcgga atgggtaagt ccggcaagag cgggaatggc
      961 tcatatcgcc tcctggatca ctacaagtac ctcaccgctt ggttcgagct gctgaacctt
     1021 ccaaagaaaa tcatctttgt gggccacgac tggggggctt gtctggcctt tcactactcc
     1081 tacgagcacc aagacaagat caaggccatc gtccatgctg agagtgtcgt ggacgtgatc
     1141 gagtcctggg acgagtggcc tgacatcgag gaggatatcg ccctgatcaa gagcgaagag
     1201 ggcgagaaaa tggtgcttga gaataacttc ttcgtcgaga ccatgctccc aagcaagatc
     1261 atgcggaaac tggagcctga ggagttcgct gcctacctgg agccattcaa ggagaagggc
     1321 gaggttagac ggcctaccct ctcctggcct cgcgagatcc ctctcgttaa gggaggcaag
     1381 cccgacgtcg tccagattgt ccgcaactac aacgcctacc ttcgggccag cgacgatctg
     1441 cctaagatgt tcatcgagtc cgaccctggg ttcttttcca acgctattgt cgagggagct
     1501 aagaagttcc ctaacaccga gttcgtgaag gtgaagggcc tccacttcag ccaggaggac
     1561 gctccagatg aaatgggtaa gtacatcaag agcttcgtgg agcgcgtgct gaagaacgag
     1621 cagtaattct aggcgatcgc tcgagagcct tcttgaagcc cccagcccta atctcaccct
     1681 ctcctccagt gtgggcttga ccaggcttgg ccttgggcta tttggactca ggtgggccct
     1741 ctgaacttgc cttaaacact caccttctag tcttggccag ccaactctgg gaatacaggg
     1801 gtgaaagggg ggaaccagtg aaaatgaaag gaagtttcag tattagatgc acttaagtta
     1861 gcctccacca ccctttcccc cttctcttag ttattgctga agagggttgg tataaaaata
     1921 attttaaaaa agccttccta cacgttagat ttgccgtacc aatctctgaa tgccccataa
     1981 ttattatttc cagtgtttgg gatgaccagg atcccaagcc tcctgctgcc acaatgttta
     2041 taaaggccaa atgatagcgg gggctaagtt ggtgcttttg agaaccaagt aaaacaaaac
     2101 gtgaccgagg agtctatttt aaagaattcg gttgaaaaaa tagatccaat cagtttatac
     2161 cctagttagt gttttgcctc acctaatagg ctgggagact gaagactcag cccgggtggg
     2221 gctgcagaaa aatgattggc cccagtcccc ttgtttgtcc cttctacagg catgaggaat
     2281 ctgggaggcc ctgagacagg gattgtgctt cattccaatc tattgcttca ccatggcctt
     2341 atgaggcagg tgagagatgt ttgaattttt ctcttccttt tagtattctt agttgttcag
     2401 ttgccaagga tccctgatcc cattttcctc tgacgtccac ctcctacccc ataggagtta
     2461 gaagttaggg tttaggcatc attttgagaa tgctgacact ttttcagggc tgtgattgag
     2521 tgagggcatg ggtaaaaata tttctttaaa agaaggatga acaattatat ttatatttca
     2581 ggttatatcc aatagtagag ttggcttttt tttttttttt tggtcatagt gggtggattt
     2641 gttgccatgt gcaccttggg gttttgtaat gacagtgcta aaaaaaaaaa agcatttttt
     2701 tttatgattt gtctctgtca cccttgtcct tgagtgctct tgctattaac gttatttgta
     2761 atttagtttg tagctcatta aaaaaatgtg cctagtttta tagcggccgc tggccgcaat
     2821 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgagga tctaaatgag
     2881 tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac gcggggggag
     2941 ggggaaggaa cgaaacactc tcattcggag gcggctcggg gtttggtctt ggtggccacg
     3001 ggcacgcaga agagcgccgc gatcctctta agcacccccc cgccctccgt ggaggcgggg
     3061 gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg cgccccagag
     3121 tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg gcgacgggct
     3181 cgctgggtcc taggctccat ggggaccgta tacgtggaca ggctctggag catccgcacg
     3241 actgcggtga tattaccgga gaccttctgc gggacgagcc gggtcacgcg gctgacgcgg
     3301 agcgtccgtt gggcgacaaa caccaggacg gggcacaggt acactatctt gtcacccgga
     3361 ggcgcgaggg actgcaggag cttcagggag tggcgcagct gcttcatccc cgtggcccgt
     3421 tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg
     3481 atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg
     3541 gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc
     3601 gagcgaccct gcagcgaccc gcttaaaagc ttggcattcc ggtactgttg gtaaagccac
     3661 catggccgat gctaagaaca ttaagaaggg ccctgctccc ttctaccctc tggaggatgg
     3721 caccgctggc gagcagctgc acaaggccat gaagaggtat gccctggtgc ctggcaccat
     3781 tgccttcacc gatgcccaca ttgaggtgga catcacctat gccgagtact tcgagatgtc
     3841 tgtgcgcctg gccgaggcca tgaagaggta cggcctgaac accaaccacc gcatcgtggt
     3901 gtgctctgag aactctctgc agttcttcat gccagtgctg ggcgccctgt tcatcggagt
     3961 ggccgtggcc cctgctaacg acatttacaa cgagcgcgag ctgctgaaca gcatgggcat
     4021 ttctcagcct accgtggtgt tcgtgtctaa gaagggcctg cagaagatcc tgaacgtgca
     4081 gaagaagctg cctatcatcc agaagatcat catcatggac tctaagaccg actaccaggg
     4141 cttccagagc atgtacacat tcgtgacatc tcatctgcct cctggcttca acgagtacga
     4201 cttcgtgcca gagtctttcg acagggacaa aaccattgcc ctgatcatga acagctctgg
     4261 gtctaccggc ctgcctaagg gcgtggccct gcctcatcgc accgcctgtg tgcgcttctc
     4321 tcacgcccgc gaccctattt tcggcaacca gatcatcccc gacaccgcta ttctgagcgt
     4381 ggtgccattc caccacggct tcggcatgtt caccaccctg ggctacctga tttgcggctt
     4441 tcgggtggtg ctgatgtacc gcttcgagga ggagctgttc ctgcgcagcc tgcaagacta
     4501 caaaattcag tctgccctgc tggtgccaac cctgttcagc ttcttcgcta agagcaccct
     4561 gatcgacaag tacgacctgt ctaacctgca cgagattgcc tctggcggcg ccccactgtc
     4621 taaggaggtg ggcgaagccg tggccaagcg ctttcatctg ccaggcatcc gccagggcta
     4681 cggcctgacc gagacaacca gcgccattct gattacccca gagggcgacg acaagcctgg
     4741 cgccgtgggc aaggtggtgc cattcttcga ggccaaggtg gtggacctgg acaccggcaa
     4801 gaccctggga gtgaaccagc gcggcgagct gtgtgtgcgc ggccctatga ttatgtccgg
     4861 ctacgtgaat aaccctgagg ccacaaacgc cctgatcgac aaggacggct ggctgcactc
     4921 tggcgacatt gcctactggg acgaggacga gcacttcttc atcgtggacc gcctgaagtc
     4981 tctgatcaag tacaagggct accaggtggc cccagccgag ctggagtcta tcctgctgca
     5041 gcaccctaac attttcgacg ccggagtggc cggcctgccc gacgacgatg ccggcgagct
     5101 gcctgccgcc gtcgtcgtgc tggaacacgg caagaccatg accgagaagg agatcgtgga
     5161 ctatgtggcc agccaggtga caaccgccaa gaagctgcgc ggcggagtgg tgttcgtgga
     5221 cgaggtgccc aagggcctga ccggcaagct ggacgcccgc aagatccgcg agatcctgat
     5281 caaggctaag aaaggcggca agatcgccgt gtaataattc tagagtcggg gcggccggcc
     5341 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     5401 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     5461 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     5521 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     5581 ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5641 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5701 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5761 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5821 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5881 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5941 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6001 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6061 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6121 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6181 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6241 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6301 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6361 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6421 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6481 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6541 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6601 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6661 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6721 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6781 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6841 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7201 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7261 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7321 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7381 gccttttgct cacatggctc gacaga
//
