LOCUS       sequence-308750-        5033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CaV1.2 5'UTR luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5033)
  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 5033)
  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_03
FEATURES             Location/Qualifiers
     source          1..5033
                     /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             894..2546
                     /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    2587..2708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2598..2617)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2652..2671
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2956..2973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3127..3715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3207..3226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3915..4706,4707..4775))
                     /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    4880..4928
                     /note="synthetic polyadenylation signal"
     misc_feature    4942..5033
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4982..5001
                     /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 gccaccaagg ttattttttc aaatggtgta
      601 gccgccggag gtgcggtgct cagttcttgg aaggggcccg gatgtactga ggatgcgtta
      661 cagtttcact cgaggaggca gtagtggaaa ggagcagttt ttggggtttg atgccataat
      721 gggaatcagg taatcgtcgg cggggaagaa gaaacgctgc agaccacggc ttcctcgaat
      781 cttgcgcgaa agccgccggc ctcggaggag ggattaatcc agacccgccg gggggtgttt
      841 tcacatttct tcctcttcgt ggctgctcct cctattaaaa ccatttttgg tccatggaag
      901 atgccaaaaa cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg
      961 gcgagcagct gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta
     1021 ccgacgcaca tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc
     1081 tggcagaagc tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg
     1141 agaatagctt gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg
     1201 ccccagctaa cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc
     1261 ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc
     1321 taccgatcat acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa
     1381 gcatgtacac cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc
     1441 ccgagagctt cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg
     1501 gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc
     1561 gcgaccccat cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat
     1621 ttcaccacgg cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg
     1681 tgctcatgta ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc
     1741 aatctgccct gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca
     1801 agtacgacct aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg
     1861 taggtgaggc cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga
     1921 cagaaacaac cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag
     1981 gcaaggtggt gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg
     2041 gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta
     2101 acaaccccga ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca
     2161 tcgcctactg ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca
     2221 aatacaaggg ctaccaggta gccccagccg aactggagag catcctgctg caacacccca
     2281 acatcttcga cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg
     2341 cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg
     2401 ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc
     2461 ctaaaggact gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca
     2521 agaagggcgg caagatcgcc gtgtaattct agagtcgggg cggccggccg cttcgagcag
     2581 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2641 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2701 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2761 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     2821 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     2881 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     2941 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3001 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3061 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3541 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc gcaaatgcta
     3901 aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc gatctgccta
     3961 tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc
     4021 ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat
     4081 ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc tgctactttg
     4141 tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg
     4201 agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg ctcgtcgttc
     4261 ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg atcacccata
     4321 ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag taagttggcc
     4381 gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt catgccatcc
     4441 gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga gtagtgtata
     4501 cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc acatagcagt
     4561 actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc aaggatcttg
     4621 ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc ttcagcatct
     4681 tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag
     4741 ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca atattattga
     4801 agcatttatc agggttacta gtacgtctct caaggataag taagtaatat taaggtacgg
     4861 gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     4921 tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa
     4981 actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tct
//
