LOCUS       V022401                 8514 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022401
VERSION     V022401
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..2045
                     /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
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             2046..2102
                     /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 porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2109..5198
                     /label="CDKL5(NM_001037343)"
                     /note="CDKL5(NM_001037343)"
                     /gene="CDKL5"
     CDS             5199..5222
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    5261..5737
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     repeat_region   5777..5917
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     rep_origin      5992..6447
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6729..6833
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6834..7694
                     /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      7865..8453
                     /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 gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1381 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1441 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1501 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1561 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     1621 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1681 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1741 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     1801 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     1861 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     1921 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     1981 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2041 acaaggccac gaacttctct ctgttaaagc aagcaggaga cgtggaagaa aaccccggtc
     2101 ctggatccat gaagattcct aacattggta atgtgatgaa taaatttgag atccttgggg
     2161 ttgtaggtga aggagcctat ggagttgtac ttaaatgcag acacaaggaa acacatgaaa
     2221 ttgtggcgat caagaaattc aaggacagtg aagaaaatga agaagtcaaa gaaacgactt
     2281 tacgagagct taaaatgctt cggactctca agcaggaaaa cattgtggag ttgaaggaag
     2341 catttcgtcg gaggggaaag ttgtacttgg tgtttgagta tgttgaaaaa aatatgctcg
     2401 aattgctgga agaaatgcca aatggagttc cacctgagaa agtaaaaagc tacatctatc
     2461 agctaatcaa ggctattcac tggtgccata agaatgatat tgtccatcga gatataaaac
     2521 cagaaaatct cttaatcagc cacaatgatg tcctaaaact gtgtgacttt ggttttgctc
     2581 gtaatctgtc agaaggcaat aatgctaatt acacagagta cgttgccacc agatggtatc
     2641 ggtccccaga actcttactt ggcgctccct atggaaagtc cgtggacatg tggtcggtgg
     2701 gctgtattct tggggagctt agcgatggac agcctttatt tcctggagaa agtgaaattg
     2761 accaactttt tactattcag aaggtgctag gaccacttcc atctgagcag atgaagcttt
     2821 tctacagtaa tcctcgcttc catgggctcc ggtttccagc tgttaaccat cctcagtcct
     2881 tggaaagaag ataccttgga attttgaata gtgttctact tgacctaatg aagaatttac
     2941 tgaagttgga cccagctgac agatacttga cagaacagtg tttgaatcac cctacatttc
     3001 aaacccagag acttctggat cgttctcctt caaggtcagc aaaaagaaaa ccttaccatg
     3061 tggaaagcag cacattgtct aatagaaacc aagccggcaa aagtactgct ttgcagtctc
     3121 accacagatc taacagcaag gacatccaga acctgagtgt aggcctgccc cgggctgacg
     3181 aaggtctccc tgccaatgaa agcttcctaa atggaaacct tgctggagct agtcttagtc
     3241 cactgcacac caaaacctac caagcaagca gccagcctgg gtctaccagc aaagatctca
     3301 ccaacaacaa cataccacac cttcttagcc caaaagaagc caagtcaaaa acagagtttg
     3361 attttaatat tgacccaaag ccttcagaag gcccagggac aaagtacctc aagtcaaaca
     3421 gcagatctca gcagaaccgc cactcattca tggaaagctc tcaaagcaaa gctgggacac
     3481 tgcagcccaa tgaaaagcag agtcggcata gctatattga cacaattccc cagtcctcta
     3541 ggagtccctc ctacaggacc aaggccaaaa gccatggggc actgagtgac tccaagtctg
     3601 tgagcaacct ttctgaagcc agggcccaaa ttgcggagcc cagtaccagt aggtacttcc
     3661 catctagctg cttagacttg aattctccca ccagcccaac ccccaccaga cacagtgaca
     3721 cgagaacttt gctcagccct tctggaagaa ataaccgaaa tgagggaacg ctggactcac
     3781 gtcgaaccac aaccagacat tctaagacga tggaggaatt gaagctgccg gagcacatgg
     3841 acagtagcca ttcccattca ctgtctgcac ctcacgaatc tttttcttat ggactgggct
     3901 acaccagccc cttttcttcc cagcaacgtc ctcataggca ttctatgtat gtgacccgtg
     3961 acaaagtgag agccaagggc ttggatggaa gcttgagcat agggcaaggg atggcagcta
     4021 gagccaacag cctgcaactc ttgtcacccc agcctggaga acagctccct ccagagatga
     4081 ctgtggcaag atcttcggtc aaagagacct ccagagaagg cacctcttcc ttccatacac
     4141 gccagaagtc tgagggtgga gtgtatcatg acccacactc tgatgatggc acagccccca
     4201 aagaaaatag acacctatac aatgatcctg tgccaaggag agttggtagc ttttacagag
     4261 tgccatctcc acgtccagac aattctttcc atgaaaataa tgtgtcaact agagtttctt
     4321 ctctaccatc agagagcagt tctggaacca accactcaaa aagacaacca gcattcgatc
     4381 catggaaaag tcctgaaaat attagtcatt cagagcaact caaggaaaaa gagaagcaag
     4441 gatttttcag gtcaatgaaa aagaaaaaga agaaatctca aacagtaccc aattccgaca
     4501 gccctgatct tctgacgttg cagaaatcca ttcattctgc tagcactcca agcagcagac
     4561 caaaggagtg gcgccccgag aagatctcag atctgcagac ccaaagccag ccattaaaat
     4621 cactgcgcaa gttgttacat ctctcttcgg cctcaaatca cccggcttcc tcagatcccc
     4681 gcttccagcc cttaacagct caacaaacca aaaattcctt ctcagaaatt cggattcacc
     4741 ccctgagcca ggcctctggc gggagcagca acatccggca ggaacccgca ccgaagggca
     4801 ggccagccct ccagctgcca gacggtggat gtgatggcag aagacagaga caccattctg
     4861 gaccccaaga tagacgcttc atgttaagga cgacagaaca acaaggagaa tacttctgct
     4921 gtggtgaccc aaagaagcct cacactccgt gcgtcccaaa ccgagccctt catcgtccaa
     4981 tctccagtcc tgctccctat ccagtactcc aggtccgagg cacttccatg tgcccgacac
     5041 tccaggtccg aggcactgat gctttcagct gcccaaccca gcaatccggg ttctctttct
     5101 tcgtgagaca cgttatgagg gaagccctga ttcacagggc ccaggtaaac caagctgcgc
     5161 tcctgacata ccatgagaat gcggcactga cgggcaagga ttacaaggac gacgatgaca
     5221 agtgaagctt gcctcgagca gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     5281 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     5341 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     5401 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     5461 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     5521 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     5581 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     5641 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     5701 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     5761 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     5821 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     5881 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     5941 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     6001 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6061 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6121 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6181 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     6241 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6301 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     6361 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6421 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     6481 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6541 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6601 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     6661 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6841 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6901 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6961 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7021 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7081 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7141 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7201 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7261 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7321 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7381 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7441 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7501 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7561 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7621 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7681 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7741 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7801 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7861 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7921 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7981 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     8041 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8101 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8161 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8221 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8281 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8341 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8401 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8461 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
