LOCUS       V036371                 8385 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036371
VERSION     V036371
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..3065
                     /label="App(NM_019288)"
                     /note="App(NM_019288)"
                     /gene="App"
     CDS             3072..3137
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3161..3749
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3815..3863
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4065..4493
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4507..4836
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4903..5697
                     /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    5873..5994
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6043..6059)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6067..6083
                     /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(6091..6121)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6136..6157
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6445..7033)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7204..8064)
                     /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(8065..8169)
                     /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 ccaccatgct gcccagcctg gcactgctcc
      781 tgctggccgc ctggacggtt cgggctctgg aggtgcccac tgatggcaat gccggtctgc
      841 tggcagaacc ccagatcgcc atgttctgtg gtaaactcaa catgcacatg aatgtgcaga
      901 atggaaaatg ggagtcagac ccatcaggga ccaaaacctg cattggcacc aaggagggaa
      961 tcctgcagta ctgccaagag gtctaccctg aactgcagat cacaaacgtg gtggaagcca
     1021 accagccagt gaccatccag aactggtgca agcggggccg caagcagtgc aagacgcaca
     1081 cccacatcgt gattccttac cggtgcctag ttggtgagtt tgtaagcgat gcccttctcg
     1141 tgcccgacaa gtgcaagttt ctacaccagg agcggatgga cgtttgtgag acccatcttc
     1201 actggcatac tgttgccaaa gagacatgca gtgagaagag cactaacttg cacgactatg
     1261 gcatgctgct gccctgtggc atcgacaagt tccgaggggt cgagttcgtg tgctgcccgt
     1321 tggcggagga gagcgacagc atcgattctg cggacgcaga ggaggacgac tccgatgtct
     1381 ggtggggtgg agcggacaca gactatgctg atggcggtga agacaaagtc gtagaagtag
     1441 ccgaagagga ggaagtggcc gatgttgagg aagaagaagc tgaggatgac gaggatgtgg
     1501 aggatgggga tgaggtggag gaggaggccg aggagcccta cgaagaggcc acagagagaa
     1561 caaccagcat tgccaccact accacaacta ccactgagtc tgtggaggag gtagtccgag
     1621 aggtgtgctc tgaacaagct gagaccgggc catgccgtgc aatgatctcc cgctggtact
     1681 ttgatgtcac tgaaggaaag tgcgccccat tcttttacgg cggatgtggc ggcaacagga
     1741 acaactttga cactgaagag tactgcatgg cggtgtgtgg cagcgtgtca tcccaaagtt
     1801 tactcaagac taccagtgaa cctcttcccc aagatccggt taaacttccc acgacggcag
     1861 ccagcacccc tgacgcagtc gacaagtacc tggagacccc cggagatgag aacgagcacg
     1921 cccatttcca gaaagccaaa gagaggttgg aagccaagca ccgagagaga atgtcccagg
     1981 tcatgagaga atgggaggag gcagaacgtc aagccaagaa tttgcccaaa gctgacaaga
     2041 aggccgttat ccagcatttc caggagaaag tggaatctct ggaacaggaa gcagccaacg
     2101 agaggcagca gcttgtagag acacacatgg ccagagttga agccatgctc aatgatcgcc
     2161 gtcgcctggc cctcgagaat tacatcaccg cactgcaggc ggtgcctcca aggcctcatc
     2221 atgtgttcaa catgctgaag aagtacgtcc gtgcagagca gaaggacaga cagcacaccc
     2281 taaagcattt tgaacatgtg cgcatggtgg accccaagaa agctgctcag atccggtccc
     2341 aggttatgac acacctccgt gtgatctacg agcgcatgaa ccagtctctg tccctgctct
     2401 acaacgtccc tgccgtggct gaggagattc aagatgaagt tgacgagctg cttcagaagg
     2461 agcagaacta ctccgacgac gtcttagcca acatgatcag tgaacccaga atcagttacg
     2521 gcaacgatgc tctcatgcct tctttgactg aaacgaagac cactgtggag ctccttcccg
     2581 tgaatggcga attcagcctg gatgatctcc aaccgtggca tccttttggg gtggactctg
     2641 tgccagccaa tacagaaaat gaagttgagc ctgtcgacgc ccgccccgct gctgaccgag
     2701 gactgaccac tcgaccaggg tctgggttga caaacatcaa gacagaagag atctcagaag
     2761 tgaagatgga tgcggagttc ggacatgatt caggcttcga agtccgccat caaaaactgg
     2821 tgttctttgc agaagatgtg ggttcaaaca aaggtgccat cattggactc atggtgggtg
     2881 gcgttgtcat agcaacagtg attgtcatca ccttggtgat gctgaagaag aaacagtaca
     2941 catccatcca tcatggcgtg gtggaggttg acgctgctgt gaccccggag gagcgccacc
     3001 tctccaagat gcagcagaat ggatatgaga atccaacata caagttcttt gagcagatgc
     3061 agaacggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3121 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3181 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3241 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3301 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3361 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3421 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3481 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3541 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3601 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3661 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3721 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3781 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3841 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3901 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3961 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4021 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4081 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4141 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4201 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4261 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4321 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4381 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4441 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4501 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4561 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4621 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4681 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4741 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4801 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4861 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4921 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4981 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5041 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5101 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5161 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5221 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5281 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5341 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5401 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5461 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5521 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5581 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5641 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5701 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5761 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5821 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5881 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5941 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6001 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6061 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6121 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6181 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6241 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6301 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6361 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6421 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6481 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6541 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6601 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6661 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6721 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6781 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6841 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6901 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6961 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7021 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7081 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7141 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7201 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7261 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7321 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7381 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7441 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7501 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7561 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7621 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7681 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7741 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7801 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7861 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7921 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7981 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8041 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8101 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8161 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8221 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8281 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8341 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
