LOCUS       V032050                 6698 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032050
VERSION     V032050
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2230
                     /label="Mapt(NM_001038609)"
                     /note="Mapt(NM_001038609)"
                     /gene="Mapt"
     CDS             2237..2260
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2298..2522
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2568..2996
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3010..3339
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3406..4200
                     /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    4374..4495
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4544..4560)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4568..4584
                     /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(4592..4622)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4637..4658
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4946..5531)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5702..6562)
                     /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(6563..6667)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggctgacc ctcgccagga
      961 gtttgacaca atggaagacc atgctggaga ttacactctg ctccaagacc aagaaggaga
     1021 catggaccat ggcttaaaag agtctccccc acagcccccc gccgatgatg gagcggagga
     1081 accagggtcg gagacctccg atgctaagag cactccaact gctgaagacg tgactgcgcc
     1141 cctagtggat gagagagctc ccgacaagca ggccgctgcc cagccccaca cggagatccc
     1201 agaaggaatt acagccgaag aagcaggcat cggagacacc ccgaaccagg aggaccaagc
     1261 cgctgggcat gtgactcaag ctcgtgtggc cagcaaagac aggacaggaa atgacgagaa
     1321 gaaagccaag ggcgctgatg gcaaaaccgg ggcgaagatc gccacacctc ggggagcagc
     1381 ctctccggcc cagaagggca cgtccaacgc caccaggatc ccggccaaga ccacgcccag
     1441 ccctaagact cctccagggt caggtgaacc accaaaatcc ggagaacgaa gcggctacag
     1501 cagccccggc tctcccggaa cgcctggcag tcgctcgcgc accccatccc taccaacacc
     1561 gcccacccgg gagcccaaga aggtggcagt ggtccgcact ccccctaagt caccatcagc
     1621 tagtaagagc cgcctgcaga ctgcccctgt gcccatgcca gacctaaaga atgtcaggtc
     1681 gaagattggc tctactgaga acctgaagca ccagccagga ggtggcaagg tgcagataat
     1741 taataagaag ctggatctta gcaacgtcca gtccaagtgt ggctcgaagg ataatatcaa
     1801 acacgtcccg ggtggaggca gtgtgcaaat agtctacaag ccggtggacc tgagcaaagt
     1861 gacctccaag tgtggctcgt tagggaacat ccatcacaag ccaggaggtg gccaggtgga
     1921 agtaaaatca gagaagctgg acttcaagga cagagtccag tcgaagattg gctccttgga
     1981 taatatcacc cacgtccctg gaggagggaa taagaagatt gaaacccaca agctgacctt
     2041 cagggagaat gccaaagcca agacagacca tggagcagaa attgtgtata agtcacccgt
     2101 ggtgtctggg gacacatctc cacggcacct cagcaatgtg tcttccacgg gcagcatcga
     2161 catggtggac tcaccacagc ttgccacact agccgatgaa gtgtctgctt ccttggccaa
     2221 gcagggtttg ctcgaggatt acaaggatga cgacgataag tagtgagggc ccgtttaaac
     2281 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2341 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2401 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2461 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2521 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2581 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2641 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2701 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2761 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2821 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2881 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2941 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3001 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3241 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3361 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3421 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3481 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3541 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3601 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3661 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3721 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3781 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3841 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3901 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3961 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4021 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4081 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4141 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4201 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4261 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4321 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4381 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4441 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4501 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4561 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4621 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4681 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4741 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4801 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4861 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4921 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5461 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5701 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5761 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5821 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5881 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5941 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6001 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6061 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6121 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6181 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6241 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6301 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6361 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6421 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6481 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6541 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6601 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6661 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
