LOCUS       sequence-234719-        6792 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 6792)
  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 6792)
  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: dre-cdk2 3'UTR sicheck2
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        81..272
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(104..124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      289..424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          508..640
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     623..642
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        685..703
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      707..716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             713..1648
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2226..2274
                     /note="synthetic polyadenylation signal"
     promoter        2283..3034
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3064..3073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3070..4722
                     /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             4029..4037
                     /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    4766..4887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4777..4796)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4831..4850
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5020..5124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5125..5193,5194..5985)
                     /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(5343..5362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6156..6744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6645..6664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctcaca tggctcgaca gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
       61 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
      121 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
      241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
      301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
      361 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
      421 ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaagctgc
      481 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
      541 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
      601 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat
      661 tacagctctt aaggctagag tacttaatac gactcactat aggctagcca ccatggcttc
      721 caaggtgtac gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg
      781 ctgcaagcaa atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc
      841 cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt
      901 cgtgcctcac atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa
      961 gtccggcaag agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc
     1021 ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc
     1081 ttgtctggcc tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc
     1141 tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat
     1201 cgccctgatc aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga
     1261 gaccatgctc ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct
     1321 ggagccattc aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat
     1381 ccctctcgtt aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta
     1441 ccttcgggcc agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc
     1501 caacgctatt gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg
     1561 cctccacttc agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt
     1621 ggagcgcgtg ctgaagaacg agcagtaatt ctaggcgatc gctcgagaac gagatcaact
     1681 ttggcaagac gggactaaaa gtttcacaca aaacttttct gaaaatgatt tcccagtttg
     1741 gaaatcgttc ggttttataa tcaatttgta cgctcgtttt cctaaagtca ctctgggttt
     1801 attttaatct gaagattatc cattgcaacc tgtcactgga aaaaaaacag ctgtgctaat
     1861 agtaagcgtg tgattaaaat gacccggacg atgtaaattt gcataaagca gacgtgatgt
     1921 tactaacata actttagttc ggagatgaca ttccatggtt ataatgctga tataacttgt
     1981 gacgttgtat tgcttatatc acacaagtac atatcgcatt gagcgcttcg tctagcctta
     2041 gactgaatga aactgtttag actccctgta tctttttgct gctttattcc aagattatat
     2101 tattataatt aaagatttta aacctaattt gcttttaaag tttcctaaaa atagttctct
     2161 ctgcatcttc atccattaaa ctcgtataac atttggttta tgttgttaca gcggccgctg
     2221 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaggatc
     2281 taaatgagtc ttcggacctc gcgggggccg cttaagcggt ggttagggtt tgtctgacgc
     2341 ggggggaggg ggaaggaacg aaacactctc attcggaggc ggctcggggt ttggtcttgg
     2401 tggccacggg cacgcagaag agcgccgcga tcctcttaag cacccccccg ccctccgtgg
     2461 aggcgggggt ttggtcggcg ggtggtaact ggcgggccgc tgactcgggc gggtcgcgcg
     2521 ccccagagtg tgaccttttc ggtctgctcg cagacccccg ggcggcgccg ccgcggcggc
     2581 gacgggctcg ctgggtccta ggctccatgg ggaccgtata cgtggacagg ctctggagca
     2641 tccgcacgac tgcggtgata ttaccggaga ccttctgcgg gacgagccgg gtcacgcggc
     2701 tgacgcggag cgtccgttgg gcgacaaaca ccaggacggg gcacaggtac actatcttgt
     2761 cacccggagg cgcgagggac tgcaggagct tcagggagtg gcgcagctgc ttcatccccg
     2821 tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg catgtcttta
     2881 gttctatgat gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga
     2941 tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat taaggtgacg cgtgtggcct
     3001 cgaacaccga gcgaccctgc agcgacccgc ttaaaagctt ggcattccgg tactgttggt
     3061 aaagccacca tggccgatgc taagaacatt aagaagggcc ctgctccctt ctaccctctg
     3121 gaggatggca ccgctggcga gcagctgcac aaggccatga agaggtatgc cctggtgcct
     3181 ggcaccattg ccttcaccga tgcccacatt gaggtggaca tcacctatgc cgagtacttc
     3241 gagatgtctg tgcgcctggc cgaggccatg aagaggtacg gcctgaacac caaccaccgc
     3301 atcgtggtgt gctctgagaa ctctctgcag ttcttcatgc cagtgctggg cgccctgttc
     3361 atcggagtgg ccgtggcccc tgctaacgac atttacaacg agcgcgagct gctgaacagc
     3421 atgggcattt ctcagcctac cgtggtgttc gtgtctaaga agggcctgca gaagatcctg
     3481 aacgtgcaga agaagctgcc tatcatccag aagatcatca tcatggactc taagaccgac
     3541 taccagggct tccagagcat gtacacattc gtgacatctc atctgcctcc tggcttcaac
     3601 gagtacgact tcgtgccaga gtctttcgac agggacaaaa ccattgccct gatcatgaac
     3661 agctctgggt ctaccggcct gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg
     3721 cgcttctctc acgcccgcga ccctattttc ggcaaccaga tcatccccga caccgctatt
     3781 ctgagcgtgg tgccattcca ccacggcttc ggcatgttca ccaccctggg ctacctgatt
     3841 tgcggctttc gggtggtgct gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg
     3901 caagactaca aaattcagtc tgccctgctg gtgccaaccc tgttcagctt cttcgctaag
     3961 agcaccctga tcgacaagta cgacctgtct aacctgcacg agattgcctc tggcggcgcc
     4021 ccactgtcta aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc
     4081 cagggctacg gcctgaccga gacaaccagc gccattctga ttaccccaga gggcgacgac
     4141 aagcctggcg ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac
     4201 accggcaaga ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt
     4261 atgtccggct acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg
     4321 ctgcactctg gcgacattgc ctactgggac gaggacgagc acttcttcat cgtggaccgc
     4381 ctgaagtctc tgatcaagta caagggctac caggtggccc cagccgagct ggagtctatc
     4441 ctgctgcagc accctaacat tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc
     4501 ggcgagctgc ctgccgccgt cgtcgtgctg gaacacggca agaccatgac cgagaaggag
     4561 atcgtggact atgtggccag ccaggtgaca accgccaaga agctgcgcgg cggagtggtg
     4621 ttcgtggacg aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag
     4681 atcctgatca aggctaagaa aggcggcaag atcgccgtgt aataattcta gagtcggggc
     4741 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     4801 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4861 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4921 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     4981 atcgataagg atccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6781 ttttgctggc ct
//
