LOCUS       V029904                 9779 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029904
VERSION     V029904
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             739..3675
                     /label="Cdkl5(NM_001421524)"
                     /note="Cdkl5(NM_001421524)"
                     /gene="Cdkl5"
     CDS             3676..3705
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3721..3777
                     /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             3778..4488
                     /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"
     misc_feature    4555..5143
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5209..5257
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5459..5887
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5901..6230
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6297..7091
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7267..7388
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7437..7453)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7461..7477
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7485..7515)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7530..7551
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7839..8427)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8598..9458)
                     /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"
     promoter        complement(9459..9563)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagccaccat gaagattcct aacattggta atgtgatgaa taaatttgag
      781 atccttgggg ttgtcggtga aggagcctat ggagttgtac ttaaatgcag acacaaggaa
      841 acacatgaaa ttgtggcaat caagaaattc aaggacagcg aagaaaatga agaagtcaag
      901 gagacgacct tacgagagct taaaatgctt cgcactctca agcaggaaaa cattgtggag
      961 ctgaaggaag cttttcgtcg gagaggaaaa ttatacctgg tgtttgaata tgttgagaag
     1021 aatatgcttg aattgctgga agaaatgcca aatggagttc cacctgaaaa agtcaagagc
     1081 tatatctacc aactaatcaa agctatccac tggtgccaca agaacgacat cgtccacaga
     1141 gatataaagc ctgaaaatct cttaatcagc cacaatgatg tcctgaaact atgtgacttt
     1201 ggttttgctc gcaatctctc agagggcaat aatgctaatt atacagagta tgtggctaca
     1261 aggtggtacc gatccccaga actgctactt ggagccccct atggaaagtc agtggacatg
     1321 tggtcagtgg gctgtattct tggagagctt agtgatggcc agcccttatt tcctggagaa
     1381 agtgaaattg accaactctt tactattcag aaggtgctag gaccacttcc atctgagcaa
     1441 atgaagctct tctacagtaa ccctcgtttc catgggctcc ggttcccagc tgttaaccat
     1501 cctcagtcat tagagagaag ataccttgga attttaaata gtgttttact tgacctaatg
     1561 aagaatttac tgaagttgga cccagctgac agatacttga cagagcagtg tttaaatcat
     1621 cctacatttc agactcagag acttttggac cggtctcctt caagatcaac aaaaaggaaa
     1681 ccttatcatg tggaaagcag caccctgtct aatagaaacc aatccaccaa gggtgctgcc
     1741 ttacagactc accacagatc caacagcaag gacattcaga acttaagtgt gggtctgccc
     1801 agggctgagg aaggcctgcc tgcaaacgaa agcttcctca atggaaacct tgctggagca
     1861 actctcagcc caatgcacac caaaacctac caagcaagca cccagcctgg ctcttccagc
     1921 aaagatctca caaacaacaa cataccacac cttctcagtc caaaagaagc caagtccaaa
     1981 acagagtttg acttcaatat tgacacaaag ccttcagaag gtccaggcac aaagtacctc
     2041 aagtccagca ccagatctca gcagaacagg cactcattca tggaaagctc ccagagcaaa
     2101 gctgggaccc tgcagcccag tgagaagcag agtcggcaca gctatattga caccattccc
     2161 cagtcctccc ggagtccatc ctacaggaca aaggccaaaa gccatggggc gttgagtgac
     2221 tccaagtctg tgagcaacct ttccgaagcc agagcccaaa tcacagagac taataccagt
     2281 agatacttcc catccagttg tttggacttg aactctccca ccagcccaac ccccaccagg
     2341 cacactgata cgagaacttt gctcagccct tccggtcgaa ataatagaaa tgaaggcact
     2401 ctggactcac ggcgaaccac cactagacac tcaaaaacca tggaggagtt gaagctgcct
     2461 gaacacatgg acagcagcca ttcgcattca ctgtctgcac ctcatgaatc attttcttat
     2521 gggctaggct acaccagccc cttctcctct cagcagcgtc ctcacaggca ttccatgtac
     2581 gtgacccggg acaaagtgag agcgaaaggc cttgatggaa gcttgagcat aggacaaggg
     2641 atggcagcca gagccaacag tttgcagctc ttatcacccc agcctggaga acagctccct
     2701 ccagagatga ctgtggcaag accttctgtt aaagagtcct cccgagaagg cgcctcttca
     2761 ttccacaccc gtcagaagtc tgagggtgga gtataccatg acccacactc tgacgacggc
     2821 acagctccca aagaaaatcg gcatctgtac aatgaccctg tcccaaggag agttggtagc
     2881 ttttatagag tgccatctcc acgaccagac aattctttcc atgaaaataa tgtgtccact
     2941 cgagtttctt ctctgccatc tgatagcagt tctggtacaa atcattcaaa aagacaacca
     3001 ggatttgatc catggaaaag tcctgaaaac attagtcatg ccgatcaact caaagaaaag
     3061 gagaagcaag gttttttcag gtcaatgaaa aagaaaaaga agaaaactca aacaacagag
     3121 tccagcaacg gcgagaatcc aagcatcaag aaatccctct tccctctttt taattcaaag
     3181 aatcatttaa agcatagttc ttctttgaaa aagctgcctg tagtcactcc acccatggtg
     3241 cccaacactg acggtcctga tctattgacc ttacagaaag ccattcattc ttctagcact
     3301 gcaagcagca gaccaaagga gtggcgacct gagaagctct cagatctaca gacccagagc
     3361 caaccactaa aatctcttcg aaagctgttg catctgtctt catcaaccaa tcatccagct
     3421 tcttcagatc ctcggttcca gcccttaaca gctcaacaag ccaaaaattc cttctcagaa
     3481 attcggattc accctctgag tcaggccact ggtgggagca gcaacatccg gcaggagcct
     3541 acaccaaaag gcagaccagc ccttcagttg ccaggatctt ccctcttaag gtacaacgga
     3601 tggaaacact caagatcaag atccagccag ccagacgaag tgatcttctt ggcatcccat
     3661 gagaaatgga aacaggagca aaagctcatt tctgaagagg acttggaatt cggaagcgga
     3721 gctactaact tcagcctgct gaagcaggct ggcgacgtgg aggagaaccc tggacctatg
     3781 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     3841 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     3901 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     3961 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     4021 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     4081 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     4141 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     4201 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     4261 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     4321 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     4381 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     4441 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaact cgagtctaga
     4501 gggcccctta agggttcgat ccctaccggt tagtaatgag tttgatatct cgacaatcaa
     4561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4681 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4741 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4801 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4861 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4921 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     4981 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5041 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5101 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     5161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5221 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5281 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5341 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5401 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     5461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5581 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     5641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5821 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5881 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     6181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     6241 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     6301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     6361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     6421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     6481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6721 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6841 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     7021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     7081 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     7141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7261 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7381 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     8041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9601 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     9661 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     9721 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
