LOCUS       sequence-406381-        3481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing GFP driven by minimal CMV promoter. This cassette 
            can be cloned into MPRA vectors to serve as a reporter gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3481)
  AUTHORS   Oliveros W, Delfosse K, Lato DF, Kiriakopulos K, Mokhtaridoost M, 
            Said A, McMurray BJ, Browning JWL, Mattioli K, Meng G, Ellis J, 
            Mital S, Mele M, Maass PG
  TITLE     Systematic characterization of regulatory variants of blood pressure
            genes.
  JOURNAL   Cell Genom. 2023 May 24;3(7):100330. doi: 
            10.1016/j.xgen.2023.100330. eCollection 2023 Jul 12.
  PUBMED    37492106
REFERENCE   2  (bases 1 to 3481)
  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: "Cell 
            Genom."; date: "2023-05-24"; volume: "3(7):100330. doi"; pages: " 
            10.1016/j.xgen.2023.100330. eCollection 2023 Jul 12"
COMMENT     Sequence Label: MPRA_minCMV+GFP
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        181..212
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      242..251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             248..946
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     polyA_signal    1035..1156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1046..1065)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1100..1119
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1404..1421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1575..2163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1655..1674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2363..3154,3155..3223))
                     /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    3328..3376
                     /note="synthetic polyadenylation signal"
     misc_feature    3390..3481
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3430..3449
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccgcttcac tgcagcaggc tgtggctctg attggctttc tggccgtcag
       61 gaacatgtcc caacatgttg agctctggca tagaagaggc tggtggctat tttgtccggt
      121 acctgagctc gctagcctcg aggatatcaa gatctggcct cggcggccaa gcttagacac
      181 tagagggtat ataatggaag ctcgacttcc agcttggcaa tccggtactg taccggtagt
      241 cgccaccatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
      301 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccgagca
      361 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct
      421 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
      481 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
      541 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
      601 cggcgacttc aaggtgatgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
      661 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgatctgga
      721 tggcagcttc acccgcacct tcagcctgcg cgacggcggc tactacagct ccgtggtgga
      781 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
      841 gttcgccttc cgccgcgtgg aggaggatca cagcaacacc gagctgggca tcgtggagta
      901 ccagcacgcc ttcaagaccc cggatgcaga tgccggtgaa gaataataat tctagacgac
      961 tcgactagat cggaagagcg tcggcctcgg cggccaagct tgtcggggcg gccggccgct
     1021 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1081 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1141 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1201 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     1261 tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     1321 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     1381 tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1441 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1501 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1741 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1981 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2041 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2221 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2281 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc
     2341 aaatgctaaa ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga
     2401 tctgcctatt tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc
     2461 gtgagggctt accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg
     2521 cccccgattt gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg
     2581 ctactttgtc cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt
     2641 cgccagtgag tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct
     2701 cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat
     2761 cacccatatt atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta
     2821 agttggccgc ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca
     2881 tgccatccgt aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt
     2941 agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac
     3001 atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa
     3061 ggatcttgcc gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt
     3121 cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg
     3181 caaagaaggg aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat
     3241 attattgaag catttatcag ggttactagt acgtctctca aggataagta agtaatatta
     3301 aggtacggga ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt
     3361 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     3421 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     3481 t
//
