LOCUS       sequence-305112-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5'UTR stem-loop luciferase reporter: 0kcal/mol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4827)
  AUTHORS   Kim JW, Yin X, Jhaldiyal A, Khan MR, Martin I, Xie Z, Perez-Rosello 
            T, Kumar M, Abalde-Atristain L, Xu J, Chen L, Eacker SM, Surmeier 
            DJ, Ingolia NT, Dawson TM, Dawson VL
  TITLE     Defects in mRNA Translation in LRRK2-Mutant hiPSC-Derived 
            Dopaminergic Neurons Lead to Dysregulated Calcium Homeostasis.
  JOURNAL   Cell Stem Cell. 2020 Oct 1;27(4):633-645.e7. doi: 
            10.1016/j.stem.2020.08.002. Epub 2020 Aug 25.
  PUBMED    32846140
REFERENCE   2  (bases 1 to 4827)
  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"; doi: 
            "10.1016/j.stem.2020.08.002"; journalName: "Cell Stem Cell"; date: 
            "2020-10-1- 1"; volume: "27"; issue: "4"; pages: "633-645.e7"
COMMENT     Sequence Label: pJWK_VD_01
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..541
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     461..479
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             688..2340
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2381..2502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2392..2411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2446..2465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2750..2767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2921..3509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3001..3020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3709..4500,4501..4569))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4674..4722
                     /note="synthetic polyadenylation signal"
     misc_feature    4736..4827
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4776..4795
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagtcttcca cggggttggg gttgcgcctt ttccaaggca
       61 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
      121 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
      181 ctacccttgt gggccccccg gcgacgcttc ctcgtccgcc cctaagtcgg gaaggttcct
      241 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
      301 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
      361 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
      421 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
      481 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
      541 aaagcttggc aatccggtac tgttggtaaa gccaccacgg aagatcacaa ctatacctaa
      601 gcatcacctc tacaactacg atcaccacta taaagatcac aactatacct aagcatcacc
      661 tctacaacta cgatcaccac tccacggatg gaagatgcca aaaacattaa gaagggccca
      721 gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag
      781 cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt
      841 acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg
      901 ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc
      961 gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag
     1021 cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa
     1081 gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc
     1141 atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat
     1201 ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc
     1261 atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg
     1321 caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc
     1381 atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc
     1441 acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag
     1501 ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta
     1561 tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag
     1621 atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc
     1681 cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc
     1741 acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct
     1801 aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc
     1861 gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc
     1921 atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac
     1981 ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca
     2041 gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc
     2101 ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa
     2161 accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag
     2221 ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac
     2281 gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgtaa
     2341 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     2401 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2461 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2521 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     2581 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     2641 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2701 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     2761 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2821 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2881 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2941 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3001 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3061 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3121 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3181 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3241 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3301 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3361 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3421 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3661 agtaaacttg gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt
     3721 gatcagtgag gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact
     3781 ccccgtcgtg tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat
     3841 gatgccgcga gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg
     3901 gagggccgag cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg
     3961 ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat
     4021 tgctactggc atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc
     4081 ccagcggtca agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt
     4141 agggcctccg atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc
     4201 agcactacac aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga
     4261 gtactcaacc aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc
     4321 gtctatacgg gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa
     4381 tcgttcttcg gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata
     4441 gcccactctt gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg
     4501 tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg
     4561 gatgctcata ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt
     4621 ctctcaagga taagtaagta atattaaggt acgggaggta ttggacaggc cgcaataaaa
     4681 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat
     4741 acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg
     4801 caagtgcagg tgccagaaca tttctct
//
