LOCUS       sequence-356628-        2887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  adding the mini luciferase and mini promoter to the MPRA lenti 
            construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2887)
  AUTHORS   Guo MG, Reynolds DL, Ang CE, Liu Y, Zhao Y, Donohue LKH, Siprashvili
            Z, Yang X, Yoo Y, Mondal S, Hong A, Kain J, Meservey L, Fabo T, 
            Elfaki I, Kellman LN, Abell NS, Pershad Y, Bayat V, Etminani P, 
            Holodniy M, Geschwind DH, Montgomery SB, Duncan LE, Urban AE, Altman
            RB, Wernig M, Khavari PA
  TITLE     Integrative analyses highlight functional regulatory variants 
            associated with neuropsychiatric diseases.
  JOURNAL   Nat Genet. 2023 Nov;55(11):1876-1891. doi: 
            10.1038/s41588-023-01533-5. Epub 2023 Oct 19.
  PUBMED    37857935
REFERENCE   2  (bases 1 to 2887)
  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: 
            "10.1038/s41588-023-01533-5"; journalName: "Nat Genet"; date: 
            "2023-11"; volume: "55"; issue: "11"; pages: "1876-1891"
COMMENT     Sequence Label: pD2_miniluc_newP7
FEATURES             Location/Qualifiers
     source          1..2887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..45)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(199..787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(279..298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(958..1749,1750..1818))
                     /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     1581..1600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1819..1923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1991..2009
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2047..2069)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2169..2188
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2382..2404
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2396..2413
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2397..2413
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2483..2514
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     primer_bind     complement(2684..2700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2684..2700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2697..2719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2708..2724
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2732..2738,2739..2756,2757..2762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2777..2798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
       61 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1921 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1981 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2041 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2101 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2161 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     2221 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     2281 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     2341 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     2401 aacgacggcc agtgaattcg agctcggtac ctgagctcct cgaggatatc aagatctggc
     2461 ctcggcggcc aagcttagac actagagggt atataatgga agctcgactt ccagcttggc
     2521 aatccggtac tgttggtaaa gacgtgtgct cttccgatct gaagatgcca aaaacattaa
     2581 gaagggccca gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa
     2641 agccatgaag cgctacgccc gctagcagct tggcgtaatc atggtcatag ctgtttcctg
     2701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2881 gaggcgg
//
