LOCUS       V036004                 8184 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036004
VERSION     V036004
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2864
                     /label="CAPN8(NM_001143962)"
                     /note="CAPN8(NM_001143962)"
                     /gene="CAPN8"
     CDS             2871..2936
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2960..3548
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3614..3662
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3864..4292
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4306..4635
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4702..5496
                     /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    5672..5793
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5842..5858)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5866..5882
                     /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(5890..5920)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5935..5956
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6244..6832)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7003..7863)
                     /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(7864..7968)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc agcccaggca gctggtgtat
      781 ctaggcagcg ggcagccact caaggtcttg gctccaacca aaacgctttg aagtacttgg
      841 gccaggattt caagaccctg aggcaacagt gcttggactc aggggtccta tttaaggacc
      901 ctgagttccc agcatgtcca tcagctttgg gctacaagga tcttggacca ggctctccgc
      961 aaactcaagg catcatctgg aagcggccca cggagttgtg tcccagccct cagtttatcg
     1021 ttggtggagc cacgcgcaca gacatttgtc agggtggtct aggtgactgc tggcttctgg
     1081 ctgccattgc ctccctgacc ctgaatgaag agctgcttta ccgggtggtc cccagggacc
     1141 aggacttcca ggagaactat gcgggaatct ttcactttca gttctggcag tacggagagt
     1201 gggtggaggt ggtcattgac gacaggctgc ccaccaagaa tggacagctg ctcttcctac
     1261 actcggaaca aggcaatgaa ttctggagtg ccctgctgga gaaagcctat gccaagctta
     1321 atggttgtta tgaggctctc gctggaggtt ccacagtgga ggggtttgag gatttcacag
     1381 gtggcatctc tgagttttat gacctgaaga aaccaccagc caatctatat cagatcatcc
     1441 ggaaggccct ctgtgcgggg tctctgctgg gctgctccat tgatgtctcc agtgcagccg
     1501 aagccgaagc catcaccagc cagaagctgg ttaagagtca tgcgtactct gtcactggag
     1561 tcgaagaggt gaatttccag ggccatccag agaagctgat cagactcagg aatccatggg
     1621 gtgaagtgga gtggtcggga gcctggagcg atgatgcacc agagtggaat cacatagacc
     1681 cccggcggaa ggaagaactg gacaagaaag ttgaggatgg agaattctgg atgtcacttt
     1741 cagatttcgt gaggcagttc tctcggttgg agatctgcaa cctgtccccg gactctctga
     1801 gtagcgagga ggtgcacaaa tggaacctgg tcctgttcaa cggccactgg acccggggct
     1861 ccacagctgg gggctgccag aactacccag ccacgtactg gaccaatccc cagttcaaaa
     1921 tccgtttgga tgaagtggat gaggaccagg aggagagcat cggtgaaccc tgctgtacag
     1981 tgctgctggg cctgatgcag aaaaatcgca ggtggcggaa gcggatagga caaggcatgc
     2041 ttagcatcgg ctatgccgtc taccaggttc ccaaggagct ggagagtcac acggacgcac
     2101 acttgggccg ggatttcttc ctggcctacc agccctcagc ccgcaccagc acctacgtca
     2161 acctgcggga ggtctctggc cgggcccggc tgccccctgg ggagtacctg gtggtgccat
     2221 ccacatttga acccttcaaa gacggcgagt tctgcttgag agtgttctca gagaagaagg
     2281 cccaggccct agaaattggg gatgtggtag ctggaaaccc atatgagcca catcccagtg
     2341 aggtggatca ggaagatgac cagttcagga ggctgtttga gaagttggca gggaaggatt
     2401 ctgagattac tgccaatgca ctcaagatac ttttgaatga ggcgttttcc aagagaacag
     2461 acataaaatt cgatggattc aacatcaaca cttgcaggga aatgatcagt ctgttggata
     2521 gcaatggaac gggcactttg ggggcggtgg aattcaagac gctctggctg aagattcaga
     2581 agtatctgga gatctattgg gaaactgatt ataaccactc gggcaccatc gatgcccacg
     2641 agatgaggac agccctcagg aaggcaggtt tcaccctcaa cagccaggtg cagcagacca
     2701 ttgccctgcg gtatgcgtgc agcaagcttg gcatcaactt tgacagcttc gtggcttgta
     2761 tgatccgcct ggagaccctc ttcaaactat tcagccttct ggacgaagac aaggatggca
     2821 tggttcagct ctctctggcc gagtggctgt gctgcgtgtt ggtcggatcc gactacaaag
     2881 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     2941 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3001 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3061 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3121 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3181 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3241 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3301 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3361 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3421 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3481 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3541 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3601 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     3661 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3721 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3781 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3841 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3901 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3961 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4021 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4081 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4141 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4201 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4261 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4381 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4441 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4501 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4561 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4621 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4681 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4741 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4801 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4861 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4921 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4981 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5041 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5101 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5161 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5221 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5281 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5341 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5401 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5461 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5521 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5581 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5641 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5761 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5821 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6421 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6661 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7981 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8041 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8101 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8161 aattgcatga agaatctgct tagg
//
