LOCUS       40924_3052        6824 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAAV-CMV-Rluc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  AUTHORS   Chen Y, Cao L, Luo C, Ditzel DA, Peter J, Sprengel R.
  TITLE     RANGE: Gene Transfer of Reversibly Controlled Polycistronic Genes
  JOURNAL   Mol Ther Nucleic Acids 2, E85 (2013)
  PUBMED    23571608
REFERENCE   2  (bases 1 to 6824)
  AUTHORS   Chen Y, Sprengel R.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2012) Molecular Neurobiology, Max Planck institute
            for medical research, Jahnstr. 29, Heidelberg, BW 69120, Germany
REFERENCE   3  (bases 1 to 6824)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6824)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Nucleic Acids 2, E85 (2013)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-NOV-2012) Molecular Neurobiology, Max Planck institute for 
            medical research, Jahnstr. 29, Heidelberg, BW 69120, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..162
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     enhancer        354..732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        733..936
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          1072..1204
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             1235..2167
                     /codon_start=1
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    2171..2224
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             2171..2224
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2240..2956
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2981..3569
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3611..3818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3828..3971
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     rep_origin      4070..4525
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4857..4961
                     /label=AmpR promoter
     CDS             4962..5819
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5993..6581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggggggg ggggggggtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
       61 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
      121 gcgcgcagag agggagtggc caactccatc actaggggtt cctagatcta ggatcacgcg
      181 ttctagacct tttgctcaca tggctcgaca gatcttcaat attggccatt agccatatta
      241 ttcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
      301 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttggcattga
      361 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      421 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      481 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      541 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      601 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      661 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
      721 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
      781 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      841 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
      901 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcact
      961 agaagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
     1021 acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc
     1081 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     1141 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     1201 acaggtgtcc actcccagtt cagaattcgg atccatgact tcgaaagttt atgatccaga
     1261 acaaaggaaa cggatgataa ctggtccgca gtggtgggcc agatgtaaac aaatgaatgt
     1321 tcttgattca tttattaatt attatgattc agaaaaacat gcagaaaatg ctgttatttt
     1381 tttacatggt aacgcggcct cttcttattt atggcgacat gttgtgccac atattgagcc
     1441 agtagcgcgg tgtattatac cagaccttat tggtatgggc aaatcaggca aatctggtaa
     1501 tggttcttat aggttacttg atcattacaa atatcttact gcatggtttg aacttcttaa
     1561 tttaccaaag aagatcattt ttgtcggcca tgattggggt gcttgtttgg catttcatta
     1621 tagctatgag catcaagata agatcaaagc aatagttcac gctgaaagtg tagtagatgt
     1681 gattgaatca tgggatgaat ggcctgatat tgaagaagat attgcgttga tcaaatctga
     1741 agaaggagaa aaaatggttt tggagaataa cttcttcgtg gaaaccatgt tgccatcaaa
     1801 aatcatgaga aagttagaac cagaagaatt tgcagcatat cttgaaccat tcaaagagaa
     1861 aggtgaagtt cgtcgtccaa cattatcatg gcctcgtgaa atcccgttag taaaaggtgg
     1921 taaacctgac gttgtacaaa ttgttaggaa ttataatgct tatctacgtg caagtgatga
     1981 tttaccaaaa atgtttattg aatcggaccc aggattcttt tccaatgcta ttgttgaagg
     2041 tgccaagaag tttcctaata ctgaatttgt caaagtaaaa ggtcttcatt tttcgcaaga
     2101 agatgcacct gatgaaatgg gaaaatatat caaatcgttc gttgagcgag ttctcaaaaa
     2161 tgaacaactc gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     2221 ccctgcaccg ggatccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2281 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2341 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     2401 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg
     2461 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2521 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2581 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2641 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     2701 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     2761 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2821 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     2881 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2941 ggacgagctg tacaagtaaa gcggccgcaa gcttatcgat aatcaacctc tggattacaa
     3001 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3061 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3121 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3181 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3241 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3301 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3361 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3421 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3481 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3541 tcggatctcc ctttgggccg cctccccgcc tgatcgatac cgtcgactag agctcgctga
     3601 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3661 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3721 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3781 ggggaggatt gggaagacaa tagcaggcat gctggggaga gatctgagga acccctagtg
     3841 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     3901 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     3961 ggagtggcca accccccccc ccccccccct gcagcctggc gtaatagcga agaggcccgc
     4021 accgatcgcc cttcccaaca gttgcgtagc ctgaatggcg aatggcgcga cgcgccctgt
     4081 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4141 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4201 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4261 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4321 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4381 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4441 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4501 aacaaaatat taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     4561 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4621 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4681 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4741 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4801 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6121 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6361 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     6421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6721 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6781 cgcctctccc cgcgcgttgg ccgattcatt aatgcaggct gcag
//
