LOCUS       sequence-234644-        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 199 mut
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        129..320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(152..172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      337..472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          556..688
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     671..690
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        733..751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      755..764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             761..1696
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASTGSWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2274..2322
                     /note="synthetic polyadenylation signal"
     promoter        2331..3082
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3112..3121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3118..4770
                     /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             4077..4085
                     /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    4814..4935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4825..4844)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4879..4898
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5068..5172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5173..5241,5242..6033)
                     /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(5391..5410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6204..6792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6693..6712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 gctcgacaga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
      121 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      421 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      481 tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
      541 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
      601 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
      661 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
      721 ggctagagta cttaatacga ctcactatag gctagccacc atggcttcca aggtgtacga
      781 ccccgagcaa cgcaaacgca tgatcactgg gcctcagtgg tgggctcgct gcaagcaaat
      841 gaacgtgctg gactccttca tcaactacta tgattccgag aagcacgccg agaacgccgt
      901 gatttttctg catggtaacg ctgcctccac aggtagctgg aggcacgtcg tgcctcacat
      961 cgagcccgtg gctagatgca tcatccctga tctgatcgga atgggtaagt ccggcaagag
     1021 cgggaatggc tcatatcgcc tcctggatca ctacaagtac ctcaccgctt ggttcgagct
     1081 gctgaacctt ccaaagaaaa tcatctttgt gggccacgac tggggggctt gtctggcctt
     1141 tcactactcc tacgagcacc aagacaagat caaggccatc gtccatgctg agagtgtcgt
     1201 ggacgtgatc gagtcctggg acgagtggcc tgacatcgag gaggatatcg ccctgatcaa
     1261 gagcgaagag ggcgagaaaa tggtgcttga gaataacttc ttcgtcgaga ccatgctccc
     1321 aagcaagatc atgcggaaac tggagcctga ggagttcgct gcctacctgg agccattcaa
     1381 ggagaagggc gaggttagac ggcctaccct ctcctggcct cgcgagatcc ctctcgttaa
     1441 gggaggcaag cccgacgtcg tccagattgt ccgcaactac aacgcctacc ttcgggccag
     1501 cgacgatctg cctaagatgt tcatcgagtc cgaccctggg ttcttttcca acgctattgt
     1561 cgagggagct aagaagttcc ctaacaccga gttcgtgaag gtgaagggcc tccacttcag
     1621 ccaggaggac gctccagatg aaatgggtaa gtacatcaag agcttcgtgg agcgcgtgct
     1681 gaagaacgag cagtaattct aggcgatcgc tcgagaacga gatcaacttt ggcaagacgg
     1741 gactaaaagt ttcacacaaa acttttctga aaatgatttc ccagtttgga aatcgttcgg
     1801 ttttataatc aatttgtacg ctcgttttcc taaagtcact ctgggtttat tttaatctga
     1861 agattatcca ttgcaacctg tgtgacccaa aaaaacagct gtgctaatag taagcgtgtg
     1921 attaaaatga cccggacgat gtaaatttgc ataaagcaga cgtgatgtta ctaacataac
     1981 tttagttcgg agatgacatt ccatggttat aatgctgata taacttgtga cgttgtattg
     2041 cttatatcac acaagtacat atcgcattga gcgcttcgtc tagccttaga ctgaatgaaa
     2101 ctgtttagac tccctgtatc tttttgctgc tttattccaa gattatatta ttataattaa
     2161 agattttaaa cctaatttgc ttttaaagtt tcctaaaaat agttctctct gcatcttcat
     2221 ccattaaact cgtataacat ttggtttatg ttgttacagc ggccgctggc cgcaataaaa
     2281 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaggatcta aatgagtctt
     2341 cggacctcgc gggggccgct taagcggtgg ttagggtttg tctgacgcgg ggggaggggg
     2401 aaggaacgaa acactctcat tcggaggcgg ctcggggttt ggtcttggtg gccacgggca
     2461 cgcagaagag cgccgcgatc ctcttaagca cccccccgcc ctccgtggag gcgggggttt
     2521 ggtcggcggg tggtaactgg cgggccgctg actcgggcgg gtcgcgcgcc ccagagtgtg
     2581 accttttcgg tctgctcgca gacccccggg cggcgccgcc gcggcggcga cgggctcgct
     2641 gggtcctagg ctccatgggg accgtatacg tggacaggct ctggagcatc cgcacgactg
     2701 cggtgatatt accggagacc ttctgcggga cgagccgggt cacgcggctg acgcggagcg
     2761 tccgttgggc gacaaacacc aggacggggc acaggtacac tatcttgtca cccggaggcg
     2821 cgagggactg caggagcttc agggagtggc gcagctgctt catccccgtg gcccgttgct
     2881 cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga
     2941 cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc
     3001 ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc
     3061 gaccctgcag cgacccgctt aaaagcttgg cattccggta ctgttggtaa agccaccatg
     3121 gccgatgcta agaacattaa gaagggccct gctcccttct accctctgga ggatggcacc
     3181 gctggcgagc agctgcacaa ggccatgaag aggtatgccc tggtgcctgg caccattgcc
     3241 ttcaccgatg cccacattga ggtggacatc acctatgccg agtacttcga gatgtctgtg
     3301 cgcctggccg aggccatgaa gaggtacggc ctgaacacca accaccgcat cgtggtgtgc
     3361 tctgagaact ctctgcagtt cttcatgcca gtgctgggcg ccctgttcat cggagtggcc
     3421 gtggcccctg ctaacgacat ttacaacgag cgcgagctgc tgaacagcat gggcatttct
     3481 cagcctaccg tggtgttcgt gtctaagaag ggcctgcaga agatcctgaa cgtgcagaag
     3541 aagctgccta tcatccagaa gatcatcatc atggactcta agaccgacta ccagggcttc
     3601 cagagcatgt acacattcgt gacatctcat ctgcctcctg gcttcaacga gtacgacttc
     3661 gtgccagagt ctttcgacag ggacaaaacc attgccctga tcatgaacag ctctgggtct
     3721 accggcctgc ctaagggcgt ggccctgcct catcgcaccg cctgtgtgcg cttctctcac
     3781 gcccgcgacc ctattttcgg caaccagatc atccccgaca ccgctattct gagcgtggtg
     3841 ccattccacc acggcttcgg catgttcacc accctgggct acctgatttg cggctttcgg
     3901 gtggtgctga tgtaccgctt cgaggaggag ctgttcctgc gcagcctgca agactacaaa
     3961 attcagtctg ccctgctggt gccaaccctg ttcagcttct tcgctaagag caccctgatc
     4021 gacaagtacg acctgtctaa cctgcacgag attgcctctg gcggcgcccc actgtctaag
     4081 gaggtgggcg aagccgtggc caagcgcttt catctgccag gcatccgcca gggctacggc
     4141 ctgaccgaga caaccagcgc cattctgatt accccagagg gcgacgacaa gcctggcgcc
     4201 gtgggcaagg tggtgccatt cttcgaggcc aaggtggtgg acctggacac cggcaagacc
     4261 ctgggagtga accagcgcgg cgagctgtgt gtgcgcggcc ctatgattat gtccggctac
     4321 gtgaataacc ctgaggccac aaacgccctg atcgacaagg acggctggct gcactctggc
     4381 gacattgcct actgggacga ggacgagcac ttcttcatcg tggaccgcct gaagtctctg
     4441 atcaagtaca agggctacca ggtggcccca gccgagctgg agtctatcct gctgcagcac
     4501 cctaacattt tcgacgccgg agtggccggc ctgcccgacg acgatgccgg cgagctgcct
     4561 gccgccgtcg tcgtgctgga acacggcaag accatgaccg agaaggagat cgtggactat
     4621 gtggccagcc aggtgacaac cgccaagaag ctgcgcggcg gagtggtgtt cgtggacgag
     4681 gtgcccaagg gcctgaccgg caagctggac gcccgcaaga tccgcgagat cctgatcaag
     4741 gctaagaaag gcggcaagat cgccgtgtaa taattctaga gtcggggcgg ccggccgctt
     4801 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4861 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4921 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4981 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     5041 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5101 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5161 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5221 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5281 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5341 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5401 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5461 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5521 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5581 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5641 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5701 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5761 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5821 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5881 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5941 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6001 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6061 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6121 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6781 agcctatgga aa
//
