LOCUS       V021358                 6922 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021358
VERSION     V021358
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             624..2216
                     /label="Uri1(NM_011274)"
                     /note="Uri1(NM_011274)"
                     /gene="Uri1"
     CDS             2217..2240
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     misc_feature    2281..2867
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             2868..3587
                     /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"
     polyA_signal    3710..3831
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3838..4293)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4320..4424
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4426..4783
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4818..5612
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5844..5891
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6220..6808
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatggagc ccccgtccga acccgagccc gagccccagc
      661 cccttgcaga ggcctcagcc gccgctccgc tgcgcgcacc agaggtggcg cggcttcgcg
      721 aggagcagga aaaggtggtc actaactgcc aagagaaaat ccagcattgg gagaaagtag
      781 acaatgacta cagtgctctt caagagaggc tgaggacctt gcctgacaag ctgtcttacg
      841 acgtcatggt accattcggc cctcttgcct tcatgccagg aaaactagtc cacactaatg
      901 aagtcactgt tttacttggc gacaactggt ttgcaaagtg ctcagccaag caggctgtgg
      961 gtttagttga gcatcggaaa gaacatgtaa gaaaaacaat agatgatttt aaaaaagtgt
     1021 tgaaaaattt tgaatccaga gttgaattca cagaagattt gcagaagatg agtgatgctg
     1081 caggtgattt tgttgacata agagaagaaa ttaaaagtga ctttgaattt aaaggaaaac
     1141 aacgaattgc tcacaaacct cattccaaac caaaaacttc agatattttt gaagcagatt
     1201 ttgaaaatgg tgtaaaaccc aagaatacat ttgatgctga tgaactgtgg gctcgacttg
     1261 aagagctgga gagacaagaa gaactgctgg gtgaacttga aagtaaacct gatactgtga
     1321 ttgcaaatgg agaagacaga gtgtcttctg aggaggaaaa ggaaggtgca gacacaggtg
     1381 tgaatgtggt gtctccagtg acagactcct ctgccgccag cagctgtaaa aggcgtgctg
     1441 gaaatgcagg actccctaat ggtcaagtca atagtctgaa ctattcagtg aatggctcaa
     1501 attcttacca cagcaacaaa gatgatgatg aagaagaaga agatgacgat gatgatgatg
     1561 atgaagatga cgataatgaa agtgaccatg ctatttcagc agataattct attccaacaa
     1621 tatatttctc tcatactgtt gaacctaaaa gggttcgaat aaatactggg aaaaatacca
     1681 ctttaaagtt cagtgagaag aaagaggaag ccaaacggaa acgaaagagt ggtgccggca
     1741 gtcacgctac ccacgagctg cctgccatca agagtcctgc tgatatttac agagtcttcg
     1801 tggatgttgt gaatggagaa tatgttcctc gcaagtcaat cctcaagtct cgcagccgag
     1861 agaacagcgt gtgcagcgac acaagtgaga gcagcgctgc cgatgtggag gaccggcggg
     1921 ggctcctgcg gagcaccagc tctgaggagg ccgtggctac cgaggccggc gggagcagtt
     1981 tggacgaact ccaagagaac caccccaaga agcctttgcc ctcaggagtg tctgaggctt
     2041 tttctggaac tgtaatagaa aaagaatttt tatcaccctc cctagcacca tactcagcca
     2101 ttgctcacca tgccctcccc accattccag agcgaaaaga agttccatca gaagtatcag
     2161 aggaacccac aaagagggtg tcaaagttca gagctgcccg attgcagcag agaagcgatt
     2221 acaaggacga cgatgacaag tagaattctg cagtcgacgg taccgcgggc ccgggatccg
     2281 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     2341 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     2401 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     2461 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     2521 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     2581 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     2641 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     2701 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     2761 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     2821 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     2881 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2941 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3001 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3061 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     3121 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     3181 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     3241 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3301 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     3361 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     3421 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3481 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3541 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     3601 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3661 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3721 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3781 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3841 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3901 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3961 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4021 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4081 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4141 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4201 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4261 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4321 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4381 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4441 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4501 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4561 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4621 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4681 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4741 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4801 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4861 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4921 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4981 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5041 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5101 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5161 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5221 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5281 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5341 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5401 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5461 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5521 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5581 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5641 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5701 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5761 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5821 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5881 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5941 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6001 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6061 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6121 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6361 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6841 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6901 taaccgtatt accgccatgc at
//
