LOCUS       Exported                6273 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  RNA interference vector psiCHECK(TM)-2, complete sequence.
ACCESSION   AY535007
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6273)
  AUTHORS   Almond BD, Vidugiriene J, Kobs G.
  TITLE     psiCHECK(TM) Vectors Technical Bulletin, TB329
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6273)
  AUTHORS   Almond BD, Vidugiriene J, Kobs G.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JAN-2004) Scientific Communications, Promega 
            Corporation, 2800 Woods Hollow Rd., Madison, WI 53711-5399, USA
REFERENCE   3  (bases 1 to 6273)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6273
                     /organism="RNA interference vector psiCHECK(TM)-2"
                     /mol_type="other DNA"
                     /db_xref="taxon:263992"
     regulatory      7..425
                     /regulatory_class="promoter"
                     /note="SV40 early enhancer/promoter"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /note="chimeric"
     regulatory      666..684
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             694..1629
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /note="derived from Renilla"
                     /protein_id="AAS38484.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    1636..1680
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    1688..1736
                     /label=polyA
                     /note="polyA"
     polyA_signal    1688..1736
                     /note="synthetic polyadenylation signal"
     regulatory      1744..2496
                     /regulatory_class="promoter"
                     /note="HSV-TK promoter"
     promoter        1745..2496
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2526..2535
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2532..4184
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /note="derived from firefly; hluc+"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3491..3499
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    4219..4440
                     /label=SV40 late polyA
                     /note="SV40 late polyA"
     polyA_signal    4228..4349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4482..4586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4587..5447
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="Ampr"
                     /protein_id="AAS38486.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5618..6206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"