LOCUS       V026052                 8197 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026052
VERSION     V026052
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             997..3717
                     /label="EIF4G2(NM_001172705)"
                     /note="EIF4G2(NM_001172705)"
                     /gene="EIF4G2"
     polyA_signal    3797..4021
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4067..4495
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4509..4838
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4905..5699
                     /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..7030)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7201..8061)
                     /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(8062..8166)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgtgg agagtgcgat tgcagaaggg
     1021 ggtgcttctc gtttcagtgc ttcttcgggc ggaggaggaa gtaggggtgc acctcagcac
     1081 tatcccaaga ctgctggcaa cagcgagttc ctggggaaaa ccccagggca aaacgctcag
     1141 aaatggattc ctgcacgaag cactagacga gatgacaact ccgcagcaaa caactccgca
     1201 aacgaaaaag aacgacatga tgcaatcttc aggaaagtaa gaggcatact aaataagctt
     1261 actcctgaaa agtttgacaa gctatgcctt gagctcctca atgtgggtgt agagtctaaa
     1321 ctcatcctta aaggggtcat actgctgatt gtggacaaag ccctagaaga gccaaagtat
     1381 agctcactgt atgctcagct atgtctgcga ttggcagaag atgcaccaaa ctttgatggc
     1441 ccagcagcag agggtcaacc aggacagaag caaagcacca cattcagacg cctcctaatt
     1501 tccaaattac aagatgaatt tgaaaaccga actagaaatg ttgatgtcta tgataagcgt
     1561 gaaaatcccc tcctccccga ggaggaggaa cagagagcca ttgctaagat caagatgttg
     1621 ggaaacatca aattcattgg agagcttggc aagcttgatc ttattcacga atctatcctt
     1681 cataagtgca tcaaaacact tttggaaaag aagaagagag tccaactcaa agatatggga
     1741 gaggatttgg agtgcctctg tcagataatg aggacagtgg gacctagatt agaccatgaa
     1801 cgagccaagt ccttaatgga tcagtacttt gcccgaatgt gctccttgat gttaagtaag
     1861 gaattgccag caaggattcg tttcctgctg caggataccg tagagttgcg agaacaccat
     1921 tgggttcctc gcaaggcttt tcttgacaat ggaccaaaga cgatcaatca aattcgtcaa
     1981 gatgcagtaa aagatctagg ggtgtttatt cctgctccta tggctcaagg gatgagaagt
     2041 gacttctttc tggagggacc gttcatgcca cccaggatga aaatggatag ggacccactt
     2101 ggaggacttg ctgatatgtt tggacaaatg ccaggtagcg gaattggtac tggtccagga
     2161 gttatccagg atagattttc acccaccatg ggacgtcatc gttcaaatca actcttcaat
     2221 ggccatgggg gacacatcat gcctcccaca caatcgcagt ttggagagat gggaggcaag
     2281 tttatgaaaa gccaggggct aagccagctc taccataacc agagtcaggg actcttatcc
     2341 cagctgcaag gacagtcgaa ggatatgcca cctcggtttt ctaagaaagg acagcttaat
     2401 gcagatgaga ttagcctgag gcctgctcag tcgttcctaa tgaataaaaa tcaagtgcca
     2461 aagcttcagc cccagataac tatgattcct cctagtgcac aaccaccacg cactcaaaca
     2521 ccacctctgg gacagacacc tcagcttggt ctcaaaacta atccaccgct tatccaggaa
     2581 aagcctgcca agaccagcaa aaagccacca ccgtcaaagg aagaactcct taaactaact
     2641 gaaactgttg tgactgaata tctaaatagt ggaaatgcaa atgaggctgt caatggtgta
     2701 agagaaatga gggctcctaa acactttctt cctgagatgt taagcaaagt aatcatcctg
     2761 tcactagata gaagcgatga agataaagaa aaagcaagtt ctttgatcag tttactcaaa
     2821 caggaaggga tagccacaag tgacaacttc atgcaggctt tcctgaatgt attggaccag
     2881 tgtcccaaac tggaggttga catccctttg gtgaaatcct atttagcaca gtttgcagct
     2941 cgtgccatca tttcagagct ggtgagcatt tcagaactag ctcaaccact agaaagtggc
     3001 acccattttc ctctcttcct actttgtctt cagcagttag ctaaattaca agatcgagaa
     3061 tggttaacag aactttttca acaaagcaag gtcaatatgc agaaaatgct cccagaaatt
     3121 gatcagaata aggaccgcat gttggagatt ttggaaggaa agggactgag tttcttattc
     3181 ccactcctca aattggagaa ggaactgttg aagcaaataa agttggatcc atcccctcaa
     3241 accatatata aatggattaa agataacatc tctcccaaac ttcatgtaga taaaggattt
     3301 gtgaacatct taatgactag cttcttacag tacatttcta gtgaagtaaa cccccccagc
     3361 gatgaaacag attcatcctc tgctccttcc aaagaacagt tagagcagga aaaacaacta
     3421 ctactatctt tcaagccagt aatgcagaaa tttcttcatg atcacgttga tctacaagtc
     3481 agtgccctgt atgctctcca ggtgcactgc tataacagca acttcccaaa aggcatgtta
     3541 cttcgctttt ttgtgcactt ctatgacatg gaaattattg aagaagaagc tttcttggct
     3601 tggaaagaag atataaccca agagtttccg ggaaaaggca aggctttgtt ccaggtgaat
     3661 cagtggctaa cctggttaga aactgctgaa gaagaagaat cagaggaaga agctgactaa
     3721 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3781 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3841 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3901 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3961 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4021 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4081 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4141 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4201 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4261 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4321 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4381 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4441 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4501 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4621 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4681 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4741 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4801 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4861 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4921 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4981 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5041 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5101 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5161 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5221 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5281 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5341 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5401 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5461 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5521 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5581 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5641 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5701 cgggactctg gggttcgaaa 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 gctttctcat agctcacgct 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 agaagaacag tatttggtat
     6901 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6961 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7201 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7261 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7321 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7381 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7441 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7501 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7561 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7621 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7681 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7741 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7801 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7861 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7921 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7981 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8041 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8161 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
