LOCUS       V028201                 8503 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028201
VERSION     V028201
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     repeat_region   1..141
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     enhancer        221..524
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        525..727
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     intron          876..1254
                     /note="fusion between human cytomegalovirus intron A and
                     human β-globin intron 2"
                     /label="chimeric intron"
     CDS             1338..2762
                     /label="Tfeb(NM_001161722)"
                     /note="Tfeb(NM_001161722)"
                     /gene="Tfeb"
     CDS             2793..3500
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEGYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             3507..3560
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             3564..5213
                     /EC_number=""
                     /codon_start=1
                     /gene="luc+"
                     /note="enhanced&nbsp;luc+ version of the luciferase gene"
                     /product="firefly luciferase"
                     /protein_id=""
                     /transl_table=1
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV*"
                     /label="luciferase"
     polyA_signal    5250..5726
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     repeat_region   5766..5906
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     rep_origin      5981..6436
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6718..6822
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6823..7683
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      7854..8442
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt cgccaccatg gcgtcacgca tcgggctgcg catgcagctc atgcgggagc
     1381 aggcccagca ggaggagcag cgagagcgca tgcagcagca ggctgtcatg cattatatgc
     1441 aacagcagca gcagcagcag cagcagctgg gtgggccccc caccccagcc atcaacaccc
     1501 ctgtccactt ccagtcgccc ccgcctgtgc ccggggaggt gctgaaggtg cagtcctacc
     1561 tggagaaccc cacctcctac cacctgcaac agtcccagca tcagaaggtt cgggagtatc
     1621 tgtctgagac ctatgggaac aagtttgctg cccacgtgag cccagcccaa ggttccccga
     1681 agcctgcccc agcagcatcc ccaggggtgc gggctggaca cgtactgtcc acctcggccg
     1741 gcaacagtgc tcccaacagt cccatggcca tgctacatat cagctccaac cccgagaaag
     1801 agtttgatga tgtcattgac aacattatgc gcctggacag cgtgctgggc tacatcaacc
     1861 ctgagatgca gatgcctaac acgctgcccc tgtctagcag ccacctgaac gtgtacagcg
     1921 gtgaccccca ggtcacagcc tccatggtgg gtgtcaccag cagctcctgc cctgccgacc
     1981 tgactcagaa gcgagagcta acagatgctg agagcagagc cctggccaag gagcggcaga
     2041 agaaagacaa tcacaaccta attgagagaa gacgcaggtt caacatcaat gaccggatca
     2101 aggagctggg aatgctgatc cccaaggcca acgacctgga cgtgcgctgg aacaaaggca
     2161 ccatcctcaa ggcctctgtg gattacatcc ggaggatgca gaaggacctg cagaagtccc
     2221 gggagctgga gaaccactcc cggcgcctgg agatgactaa caagcagctc tggctccgca
     2281 tccaggagct ggagatgcag gcacgcgtgc acggcctccc caccacctcg ccgtcgggtg
     2341 tgaatatggc cgagctggcc cagcaggtgg tgaagcaaga gttgcccagt gaggatggcc
     2401 caggggaggc gctgatgctg gggcctgagg tccctgagcc tgagcaaatg ccggctcttc
     2461 ctccccaggc tccgctgccc tcggccgccc agccacagtc tccgttccat cacctggact
     2521 tcagccatgg cctgagcttt gggggtgggg gcgacgaggg gcccacaggt taccccgata
     2581 ccctggggac agagcacggc tccccattcc ccaacctgtc caagaaggat ctggacttaa
     2641 tgctcctaga tgactccctg ctccccctgg cctctgaccc cctcttttct accatgtctc
     2701 ctgaggcctc caaggccagc agccgccgga gcagcttcag catggaggag ggtgatgttc
     2761 tgggtggagg cggatcagga ggcggtggat ccatggtgag caagggcgag gaggataaca
     2821 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     2881 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2941 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     3001 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     3061 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     3121 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     3181 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     3241 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     3301 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     3361 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     3421 acgagggcta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     3481 gcatggacga gctgtacaag ccccgggagg gcagaggaag tcttctaaca tgcggtgacg
     3541 tggaggagaa tcccggccct cgagaagacg ccaaaaacat aaagaaaggc ccggcgccat
     3601 tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg
     3661 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg
     3721 ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
     3781 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     3841 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     3901 tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc
     3961 aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt
     4021 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     4081 ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac
     4141 tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa
     4201 ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     4261 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     4321 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc
     4381 tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct
     4441 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     4501 ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc
     4561 caggtatcag gcaaggatat gggctcactg agactacatc agctattctg attacgcccg
     4621 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     4681 tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag
     4741 gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     4801 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     4861 tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat
     4921 tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg
     4981 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     5041 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     5101 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     5161 aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg taaaagcttg
     5221 cctcgagcag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     5281 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     5341 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     5401 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     5461 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     5521 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     5581 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     5641 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     5701 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     5761 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     5821 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     5881 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     5941 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     6001 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6061 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6121 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6181 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6241 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6301 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     6361 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6421 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6481 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6541 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6601 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6661 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6721 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6781 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6841 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6901 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6961 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7021 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7081 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7141 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7201 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7261 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7321 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7381 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7441 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7501 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7561 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7621 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7681 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7741 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7801 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7861 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7921 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7981 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     8041 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8101 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8161 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8221 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8281 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8341 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8401 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8461 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
