LOCUS       Exported                9258 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pREX1LC, complete sequence.
ACCESSION   AF383623
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9258)
  AUTHORS   Murakawa M, Freeman MW.
  TITLE     Adeno-associated virus 2 (AAV)-based plasmid vector system that can 
            mediate auto-excision of AAV terminal motifs via auto-regulated 
            transient Rep78 expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9258)
  AUTHORS   Murakawa M, Freeman MW.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2001) Lipid Metabolism Unit, Massachusetts General
            Hospital, Jackson 1328, Boston, MA 02114-2694, USA
REFERENCE   3  (bases 1 to 9258)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9258
                     /organism="Cloning vector pREX1LC"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:162323"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(673..1374,1728..2891)
                     /codon_start=1
                     /product="adeno-associated virus 2 Rep78 protein"
                     /label=adeno-associated virus 2 Rep78 protein
                     /note="major coat protein A"
                     /protein_id="AAK63810.1"
                     /translation="MPGFYEIVIKVPSDLDGHLPGISDSFVNWVAEKEWELPPDSDMDL
                     NLIEQAPLTVAEKLQRDFLTEWRRVSKAPEALFFVQFEKGESYFHMHVLVETTGVKSMV
                     LGRFLSQIREKLIQRIYRGIEPTLPNWFAVTKTRNGAGGGNKVVDECYIPNYLLPKTQP
                     ELQWAWTNMEQYLSACLNLTERKRLVAQHLTHVSQTQEQNKENQNPNSDAPVIRSKTSA
                     RYMELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAGKIMSLTKTAP
                     DYLVGQQPVEDISSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGK
                     TNIAEAIAHTVPFYGCVNWTNENFPFNDCVDKMVIWWEEGKMTAKVVESAKAILGGSKV
                     RVDQKCKSSAQIDPTPVIVTSNTNMCAVIDGNSTTFEHQQPLQDRMFKFELTRRLDHDF
                     GKVTKQEVKDFFRWAKDHVVEVEHEFYVKKGGAKKRPAPSDADISEPKRVRESVAQPST
                     SDAEASINYADRYQNKCSRHVGMNLMLFPCRQCERMNQNSNICFTHGQKDCLECFPVSE
                     SQPVSVVKKAYQKLCYIHHIMGKVPDACTACDLVNVDLDDCIFEQ"
     intron          1375..1727
                     /note="artificial"
     repeat_region   1421..1565
                     /label=adeno-associated virus 5 inverted terminal repeat
                     /note="adeno-associated virus 5 inverted terminal repeat"
     misc_feature    1462..1503
                     /label=flip oriented DNA
                     /note="flip oriented DNA"
     promoter        3352..4533
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3582..4524
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      4546..4555
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4552..5271
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorescent protein"
                     /label=GFP
                     /note="derived from Renilla reniformis"
                     /protein_id="AAK63811.1"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     gene            4552..5271
                     /gene="GFP"
                     /label=GFP
     polyA_signal    5385..5506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5612..5756
                     /label=adeno-associated virus 3 inverted terminal repeat
                     /note="adeno-associated virus 3 inverted terminal repeat"
     misc_feature    5673..5715
                     /label=flop oriented DNA
                     /note="flop oriented DNA"
     CDS             complement(5771..6928)
                     /codon_start=1
                     /product="AU1-tagged herpes simplex virus-thymidine kinase"
                     /label=AU1-tagged herpes simplex virus-thymidine kinase
                     /note="HSV-TK"
                     /protein_id="AAK63812.1"
                     /translation="MGDTYRYINMASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQE
                     ATEVRLEQKMPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASE
                     TIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPP
                     PALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRH
                     IDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGA
                     EPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQ
                     SPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     CDS             complement(5771..6901)
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     misc_feature    complement(6905..6922)
                     /note="derived from bovine papilloma virus type 1; Region: 
                     AU1 epitope"
     regulatory      6925..6934
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7152..7343
                     /codon_start=1
                     /gene="synrop"
                     /product="ROP protein"
                     /label=synrop
                     /note="similar to the protein encoded by GenBank Accession 
                     Number J01749"
                     /protein_id="AAK63813.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            7152..7343
                     /gene="synrop"
                     /label=synrop
     misc_feature    7395..9237
                     /label=derived from cloning vector pBR322
                     /note="derived from cloning vector pBR322"
     rep_origin      complement(7413..8001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8172..9032)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="similar to the protein encoded by GenBank Accession 
                     Number AAB59737; confers ampicillin resistance"
                     /protein_id="AAK63814.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(8172..9032)
                     /gene="bla"
                     /label=bla
     promoter        complement(9033..9137)
                     /gene="bla"
                     /label=AmpR promoter