LOCUS       V021708                 8411 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021708
VERSION     V021708
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             872..3181
                     /label="APP(NM_000484)"
                     /note="APP(NM_000484)"
                     /gene="APP"
     CDS             3197..3913
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3920..3937
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3978..4202
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4248..4676
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4690..5019
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5086..5880
                     /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    6054..6175
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6224..6240)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6248..6264
                     /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(6272..6302)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6317..6338
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6626..7211)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7382..8242)
                     /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(8243..8347)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccac catgctgccc ggtttggcac tgctcctgct
      901 ggccgcctgg acggctcggg cgctggaggt acccactgat ggtaatgctg gcctgctggc
      961 tgaaccccag attgccatgt tctgtggcag actgaacatg cacatgaatg tccagaatgg
     1021 gaagtgggat tcagatccat cagggaccaa aacctgcatt gataccaagg aaggcatcct
     1081 gcagtattgc caagaagtct accctgaact gcagatcacc aatgtggtag aagccaacca
     1141 accagtgacc atccagaact ggtgcaagcg gggccgcaag cagtgcaaga cccatcccca
     1201 ctttgtgatt ccctaccgct gcttagttgg tgagtttgta agtgatgccc ttctcgttcc
     1261 tgacaagtgc aaattcttac accaggagag gatggatgtt tgcgaaactc atcttcactg
     1321 gcacaccgtc gccaaagaga catgcagtga gaagagtacc aacttgcatg actacggcat
     1381 gttgctgccc tgcggaattg acaagttccg aggggtagag tttgtgtgtt gcccactggc
     1441 tgaagaaagt gacaatgtgg attctgctga tgcggaggag gatgactcgg atgtctggtg
     1501 gggcggagca gacacagact atgcagatgg gagtgaagac aaagtagtag aagtagcaga
     1561 ggaggaagaa gtggctgagg tggaagaaga agaagccgat gatgacgagg acgatgagga
     1621 tggtgatgag gtagaggaag aggctgagga accctacgaa gaagccacag agagaaccac
     1681 cagcattgcc accaccacca ccaccaccac agagtctgtg gaagaggtgg ttcgagaggt
     1741 gtgctctgaa caagccgaga cggggccgtg ccgagcaatg atctcccgct ggtactttga
     1801 tgtgactgaa gggaagtgtg ccccattctt ttacggcgga tgtggcggca accggaacaa
     1861 ctttgacaca gaagagtact gcatggccgt gtgtggcagc gccatgtccc aaagtttact
     1921 caagactacc caggaacctc ttgcccgaga tcctgttaaa cttcctacaa cagcagccag
     1981 tacccctgat gccgttgaca agtatctcga gacacctggg gatgagaatg aacatgccca
     2041 tttccagaaa gccaaagaga ggcttgaggc caagcaccga gagagaatgt cccaggtcat
     2101 gagagaatgg gaagaggcag aacgtcaagc aaagaacttg cctaaagctg ataagaaggc
     2161 agttatccag catttccagg agaaagtgga atctttggaa caggaagcag ccaacgagag
     2221 acagcagctg gtggagacac acatggccag agtggaagcc atgctcaatg accgccgccg
     2281 cctggccctg gagaactaca tcaccgctct gcaggctgtt cctcctcggc ctcgtcacgt
     2341 gttcaatatg ctaaagaagt atgtccgcgc agaacagaag gacagacagc acaccctaaa
     2401 gcatttcgag catgtgcgca tggtggatcc caagaaagcc gctcagatcc ggtcccaggt
     2461 tatgacacac ctccgtgtga tttatgagcg catgaatcag tctctctccc tgctctacaa
     2521 cgtgcctgca gtggccgagg agattcagga tgaagttgat gagctgcttc agaaagagca
     2581 aaactattca gatgacgtct tggccaacat gattagtgaa ccaaggatca gttacggaaa
     2641 cgatgctctc atgccatctt tgaccgaaac gaaaaccacc gtggagctcc ttcccgtgaa
     2701 tggagagttc agcctggacg atctccagcc gtggcattct tttggggctg actctgtgcc
     2761 agccaacaca gaaaacgaag ttgagcctgt tgatgcccgc cctgctgccg accgaggact
     2821 gaccactcga ccaggttctg ggttgacaaa tatcaagacg gaggagatct ctgaagtgaa
     2881 gatggatgca gaattccgac atgactcagg atatgaagtt catcatcaaa aattggtgtt
     2941 ctttgcagaa gatgtgggtt caaacaaagg tgcaatcatt ggactcatgg tgggcggtgt
     3001 tgtcatagcg acagtgatcg tcatcacctt ggtgatgctg aagaagaaac agtacacatc
     3061 cattcatcat ggtgtggtgg aggttgacgc cgctgtcacc ccagaggagc gccacctgtc
     3121 caagatgcag cagaacggct acgaaaatcc aacctacaag ttctttgagc agatgcagaa
     3181 cggaggaggt ggatccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3241 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3301 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3361 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3421 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3481 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3541 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3601 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3661 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3721 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3781 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3841 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3901 cgagctgtac aagctcgagc accatcacca tcaccattag tctagagggc ccgtttaaac
     3961 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4021 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4081 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4141 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4201 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4441 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4561 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4681 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4741 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4861 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4921 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4981 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5041 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5101 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5161 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5221 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5281 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5341 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5401 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5461 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5521 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5581 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5641 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5701 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5761 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5821 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5881 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5941 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6001 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6181 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6241 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6301 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6361 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7141 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7201 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7261 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7321 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8341 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     8401 tcccctatgg t
//
