LOCUS       MPRA_minCMV+GFP         3481 bp    DNA     circular SYN 26-JUN-2023
DEFINITION  MPRA with Minimal Promoter + GFP + Positive Control Oligo.
ACCESSION   -
KEYWORDS    .
SOURCE      Firefly luciferase reporter vector pGL4.23[luc2/minP]
  ORGANISM  Artificial
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4283)
  AUTHORS   Swanson,B., Kopish,K. and McNamara,B.
  TITLE     pGL4 Luciferase reporter Vectors Technical Manual, TM259
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4283)
  AUTHORS   Swanson,B., Kopish,K. and McNamara,B.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-AUG-2006) Scientific Communications, Promega
            Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA
FEATURES             Location/Qualifiers
     primer_bind     7..22
                     /Sequence="ACTGGCCGCTTCACTG"
                     /Hairpin_Tm="None"
                     /Tm="56.6"
                     /Self_Dimer_Tm="None"
                     /%GC="62.5"
                     /Color="#1199ff"
                     /modified_by="User"
                     /label="Universal PCR primer site"
     misc_feature    23..117
                     /Color="#ff9911"
                     /created_by="User"
                     /label
     motif           63..78
                     /Color="#ff17e1"
                     /created_by="User"
                     /label="P53 MOTIF"
     unsure          118..123
                     /created_by="User"
                     /label="KpnI Site"
     promoter        182..212
                     /created_by="User"
                     /label="Minimal Promoter"
     ORF             248..946
                     /created_by="User"
                     /Color="#99ff99"
                     /modified_by="User"
                     /label="GFP"
     primer_bind     432..451
                     /Sequence="TGATGGGCTACGGCTTCTAC"
                     /Hairpin_Tm="38.7"
                     /Tm="59.2"
                     /Self_Dimer_Tm="None"
                     /%GC="55.0"
                     /created_by="primer3"
                     /label="tGFP-qF1"
     misc_feature    complement(605..624)
                     /Sequence="GTGCCCATCACCTTGAAGTC"
                     /Hairpin_Tm="39.8"
                     /Tm="58.8"
                     /Self_Dimer_Tm="None"
                     /%GC="55.0"
                     /created_by="primer3"
                     /label="tGFP-qR1"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     636..655
                     /Sequence="AGGACAGCGTGATCTTCACC"
                     /Hairpin_Tm="46.3"
                     /Tm="59.8"
                     /Self_Dimer_Tm="None"
                     /%GC="55.0"
                     /created_by="primer3"
                     /label="tGFP-qF2"
     misc_feature    complement(780..799)
                     /Sequence="CTTGAAGTGCATGTGGCTGT"
                     /Hairpin_Tm="45.2"
                     /Tm="59.0"
                     /Self_Dimer_Tm="None"
                     /%GC="50.0"
                     /created_by="primer3"
                     /label="tGFP-qR2"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     851..867
                     /Sequence="CGCCGCGTGGAGGAGGA"
                     /Hairpin_Tm="37.5"
                     /Tm="64.3"
                     /Self_Dimer_Tm="5.7"
                     /%GC="76.5"
                     /created_by="primer3"
                     /label="LibrarySeqPrimer"
     unsure          951..956
                     /created_by="User"
                     /label="XbaI Site"
     misc_RNA        957..966
                     /Color="#000000"
                     /created_by="User"
                     /label="Barcode"
     misc_feature    complement(967..983)
                     /Sequence="CGACGCTCTTCCGATCT"
                     /Hairpin_Tm="None"
                     /Tm="56.0"
                     /Self_Dimer_Tm="None"
                     /%GC="58.8"
                     /Color="#1199ff"
                     /modified_by="User"
                     /label="Universal PCR primer site"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1014..1030)
                     /Sequence="GTCTGCTCGAAGCGGCC"
                     /Hairpin_Tm="49.0"
                     /Tm="60.9"
                     /Self_Dimer_Tm="5.4"
                     /%GC="70.6"
                     /modified_by="User"
                     /label="Gene Specific RT Primer"
                     /note="Geneious type: primer_bind_reverse"
     polyA_signal    1026..1247
                     /note="SV40 late poly(A) signal"
     rep_origin      1572
                     /note="ColE1-derived plasmid replication origin"
     CDS             complement(2363..3223)
                     /gene="Amp(r)"
                     /codon_start=1
                     /transl_table=11
                     /product="synthetic beta-lactamase"
                     /protein_id="ABI94440.1"
                     /db_xref="GI:115342876"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(2363..3223)
                     /gene="Amp(r)"
     polyA_signal    3328..3481
                     /note="poly(A) signal/transcriptional pause site"
     primer_bind     3430..3449
                     /note="RVprimer3 binding region"
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
//
