LOCUS       sequence-308747-        4823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5'UTR stem-loop luciferase reporter: -30kcal/mol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4823)
  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 4823)
  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_02
FEATURES             Location/Qualifiers
     source          1..4823
                     /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             684..2336
                     /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    2377..2498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2388..2407)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2442..2461
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2746..2763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2917..3505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2997..3016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3705..4496,4497..4565))
                     /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    4670..4718
                     /note="synthetic polyadenylation signal"
     misc_feature    4732..4823
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4772..4791
                     /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 taaggtggaa tttgtaacgt agaggcttgt
      661 attgagtact agtacgatct accatggaag atgccaaaaa cattaagaag ggcccagcgc
      721 cattctaccc actcgaagac gggaccgccg gcgagcagct gcacaaagcc atgaagcgct
      781 acgccctggt gcccggcacc atcgccttta ccgacgcaca tatcgaggtg gacattacct
      841 acgccgagta cttcgagatg agcgttcggc tggcagaagc tatgaagcgc tatgggctga
      901 atacaaacca tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt
      961 tgggtgccct gttcatcggt gtggctgtgg ccccagctaa cgacatctac aacgagcgcg
     1021 agctgctgaa cagcatgggc atcagccagc ccaccgtcgt attcgtgagc aagaaagggc
     1081 tgcaaaagat cctcaacgtg caaaagaagc taccgatcat acaaaagatc atcatcatgg
     1141 atagcaagac cgactaccag ggcttccaaa gcatgtacac cttcgtgact tcccatttgc
     1201 cacccggctt caacgagtac gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg
     1261 ccctgatcat gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc
     1321 gcaccgcttg tgtccgattc agtcatgccc gcgaccccat cttcggcaac cagatcatcc
     1381 ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc
     1441 tgggctactt gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag gaggagctat
     1501 tcttgcgcag cttgcaagac tataagattc aatctgccct gctggtgccc acactattta
     1561 gcttcttcgc taagagcact ctcatcgaca agtacgacct aagcaacttg cacgagatcg
     1621 ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa cgcttccacc
     1681 taccaggcat ccgccagggc tacggcctga cagaaacaac cagcgccatt ctgatcaccc
     1741 ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg
     1801 tggtggactt ggacaccggt aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc
     1861 gtggccccat gatcatgagc ggctacgtta acaaccccga ggctacaaac gctctcatcg
     1921 acaaggacgg ctggctgcac agcggcgaca tcgcctactg ggacgaggac gagcacttct
     1981 tcatcgtgga ccggctgaag agcctgatca aatacaaggg ctaccaggta gccccagccg
     2041 aactggagag catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc
     2101 ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca
     2161 tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc
     2221 gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc
     2281 gcaagatccg cgagattctc attaaggcca agaagggcgg caagatcgcc gtgtaattct
     2341 agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat gagtttggac
     2401 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2461 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2521 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     2581 aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt caacccagtc
     2641 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     2701 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
     2761 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3661 aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca gtgcttgatc
     3721 agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc ctgactcccc
     3781 gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc agcaatgatg
     3841 ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg
     3901 gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat gagctgctgt
     3961 cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt ggccattgct
     4021 actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc tggttcccag
     4081 cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag ctccttaggg
     4141 cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt aatggcagca
     4201 ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac cggcgagtac
     4261 tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg cccggcgtct
     4321 atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat cgggaatcgt
     4381 tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc gatatagccc
     4441 actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc ggggtgtgca
     4501 aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg
     4561 ctcatactcg tcctttttca atattattga agcatttatc agggttacta gtacgtctct
     4621 caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca ataaaatatc
     4681 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     4741 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     4801 tgcaggtgcc agaacatttc tct
//
