LOCUS       V021111                 6544 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021111
VERSION     V021111
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"
     regulatory      618..627
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             627..650
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             651..2456
                     /label="GLS2(NM_013267)"
                     /note="GLS2(NM_013267)"
                     /gene="GLS2"
     CDS             2490..3209
                     /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    3332..3453
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3460..3915)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3942..4046
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4048..4405
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4440..5234
                     /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    5466..5513
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5842..6430
                     /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 accatggatt acaaggacga cgatgacaag atgcgctcca
      661 tgaaggctct gcagaaggcc ctgagccggg ctggcagtca ctgcgggcga ggaggctggg
      721 gtcacccgag ccggagcccc ctccttggcg ggggcgtccg gcaccacctc agtgaggccg
      781 cggcgcaggg cagagagacg ccacacagcc accagccgca gcaccaggat catgattcat
      841 cagaaagtgg catgctgtcc cgcctgggtg atttgctctt ttacactatt gctgaaggac
      901 aggaacgaat ccctatccac aagttcacca ctgcactaaa ggccactgga ctgcagacat
      961 cagatcctcg gctccgagac tgcatgagcg agatgcaccg cgtggtccaa gagtccagta
     1021 gtggtggcct cttggaccga gatctcttcc gaaagtgtgt gagcagcaac attgtgctcc
     1081 tgacccaggc attccgaaag aagtttgtca ttcctgattt tgaggagttc acgggccatg
     1141 tggatcgcat ctttgaggat gtcaaagagc tcactggagg caaagtggca gcctacatcc
     1201 ctcagctggc caagtcaaac ccagacctgt ggggtgtctc cctgtgcact gtggatggtc
     1261 aacggcactc tgtgggccac acaaagatcc ccttctgcct gcagtcctgt gtgaagcccc
     1321 tcacctatgc catctccata agcaccctag gcactgacta cgtgcacaag tttgtgggca
     1381 aagagccaag tggcctgcgc tacaacaagc tctccctcaa tgaggaagga atcccccata
     1441 accccatggt caatgctggt gccattgttg tcagctccct gatcaagatg gactgtaaca
     1501 aagcagagaa gtttgatttt gtgttgcagt atctcaacaa aatggctggg aatgaataca
     1561 tgggtttcag caatgccaca ttccagtcag agaaggaaac aggggatcgg aattatgcca
     1621 tcggctatta tctcaaggaa aagaagtgct ttcctaaggg ggtggacatg atggctgccc
     1681 ttgatctcta cttccagctg tgttctgtgg aggtcacttg tgaatcaggc agtgtcatgg
     1741 cagccaccct cgccaacggt gggatctgcc ccatcacagg cgagagtgtg ctgagtgctg
     1801 aagcagtgcg caacaccctc agcctcatgc attcctgcgg catgtatgac ttctctggcc
     1861 agtttgcctt ccacgtgggc ctgccagcca agtcagctgt atcaggagcc atcctcctgg
     1921 tggtacccaa tgtcatggga atgatgtgcc tgtcaccccc attggacaag ctggggaaca
     1981 gccatagggg gaccagcttc tgccagaagt tggtgtctct cttcaatttc cacaactatg
     2041 acaacctgag gcactgtgct cggaagttag acccacggcg tgaaggggca gaaattcgga
     2101 acaagactgt ggtcaacctg ttatttgctg cctatagtgg cgatgtctca gctcttcgaa
     2161 ggtttgcctt gtcagccatg gatatggaac agaaagacta tgactcgcgc acagctctgc
     2221 atgttgctgc agctgaagga cacatcgaag ttgttaaatt cctgatcgag gcttgcaaag
     2281 tgaatccttt tgccaaggac aggtggggca acattcccct ggatgatgct gtgcagttca
     2341 accatctgga ggtggtcaaa ctgcttcaag attaccagga ctcctacaca ctctctgaaa
     2401 ctcaggctga ggcagcagct gaggccctgt ccaaagagaa cttagaaagc atggtaggag
     2461 gagggggaag cgatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2521 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2581 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2641 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2701 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2761 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2821 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2881 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2941 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3001 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3061 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3121 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3181 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     3241 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3301 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3361 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3421 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     3481 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3541 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3601 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3661 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3721 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3781 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3841 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3901 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3961 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4021 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4081 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4141 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4201 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4261 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4321 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4381 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4441 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4501 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4561 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4621 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4681 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4741 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4801 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4861 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4921 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4981 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5041 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5101 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5161 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5221 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5281 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5341 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5401 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5461 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5521 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5581 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5641 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5701 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5761 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5821 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5881 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5941 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6001 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6061 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6481 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6541 gcat
//
