LOCUS       V036037                 8247 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036037
VERSION     V036037
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..2927
                     /label="Eef2k(NM_012947)"
                     /note="Eef2k(NM_012947)"
                     /gene="Eef2k"
     CDS             2934..2999
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3023..3611
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3677..3725
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3927..4355
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4369..4698
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4765..5559
                     /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    5735..5856
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5905..5921)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5929..5945
                     /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(5953..5983)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5998..6019
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6307..6895)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7066..7926)
                     /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(7927..8031)
                     /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 ccaccatggc agacgaagac ctcatcttcc
      781 gcctggaagg tgttgacggt ggcgggtcct ccggagctgg ccgccatggg gactctgaca
      841 cagacagtga cgatgatgag ggctacttca tctgccccat cactgatgac cacatgtcca
      901 accagaatgt caactccaaa ggccagggct actacaacaa cctactgaaa acggagtgtg
      961 gctccacagg gtcaccagcc agctccttcc acttcaagga ggcctggaag catgcgatcg
     1021 agaaggccaa gcacatgccc gacccctggg ctgaattcca tctggaggac atcgccacag
     1081 agcatgctac tcggcacagg tacaatgctg tcaccgggga gtggctgaaa gacgaggttc
     1141 tgatcaagat ggcatctcag cccttcggcc gtggagcaat gagggagtgc ttcaggacga
     1201 agaaactctc caacttcttg catgcccagc attggaaggg ggcctccaac tacgtggcaa
     1261 aacgctacct cgagccagtg gacaggagcg tgtactttga ggatgtgcag ctccagatgg
     1321 aagcgaagct ctggggggag gagtacaacc ggcacaagcc ccccaagcag gtagatatca
     1381 tgcagatgtg catcattgag ctaaaggaca gacaaggaca gcccctcttc cacctggagc
     1441 actacattga gggcaagtac atcaagtata attccaactc aggctttgtc cgtgatgaca
     1501 acatcagact aacaccacag gccttcagcc atttcacatt tgaacgttct ggtcatcagc
     1561 tgattgtagt ggacatccag ggtgtgggcg acctttatac tgacccacag atccacactg
     1621 agaaaggcac tgactttgga gatggtaacc ttggtgtccg gggaatggct ctcttcttct
     1681 actctcatgc ctgcaaccgt atttgccaga gcatgggcct tgcgcccttt gacctctccc
     1741 cacgggagca ggatgccgtg aaccagagca ccaagctgtt gcaatcagcc aagaccatct
     1801 tgagggggac agaggagaag tgtgggagtc cccgcataag gacactctct ggcagccggc
     1861 cccccttgct ccttcgcctg tcagagaact ctggggatga gaacatgagt gatgtgacct
     1921 ttgactctct gccttcctcc ccgtcttcag ccacaccaca cagccagaag ttggaccacc
     1981 tccattggcc agtgtttggt gacctcgata acatgggccc tagagaccat gaccgcatgg
     2041 acaatcaccg ggactctgag aatagtgggg acagtgggta tccaagcgag aagcgaagtg
     2101 acctggatga tcctgagccc cgagaacacg gccactccaa tggcaaccga aggcctgaat
     2161 ctgatgagga cagcctgggc agctctggac gggtctgtgt ggagacgtgg aacctgctca
     2221 atccctcccg cctgcacctg cccaggccct cggccgtggc cctagaagtg caaaggttaa
     2281 atgctctgga ccttggaagg aaaatcggga agtctgtttt ggggaaggtc catctggcca
     2341 tggtacggta ccacgagggc gggcgcttct gcgagaagga tgaggaatgg gatcaagagt
     2401 ctgccatctt ccatctggag catgcagccg acctgggaga actggaggcc atcgtgggcc
     2461 taggcctcat gtactctcag ctgccccacc acatcctggc tgatgtctct ctggaggaga
     2521 cagaagagaa caagacaaaa ggctttgatt acctcctgaa ggccgcagaa gctggtgaca
     2581 ggcagtccat gattttagtg gctcgagctt ttgacaccgg cctgaacctc agcccagaca
     2641 ggtgtcaaga ctggtcggaa gccttacact ggtacaacac ggccctggag acgacggact
     2701 gcgatgaagg cggggagtat gacggaatac aggacgagcc ccagtatgca ctgctggcca
     2761 gggaggcaga gatgttgctc accggtggat ttggactgga caagaacccc caaagatcag
     2821 gagacttgta cacccaggca gccgaggcag caatggaagc catgaagggc cggctagcca
     2881 accagtacta tgagaaggcg gaagaggcct gggcgcaaat ggaggaagga tccgactaca
     2941 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3001 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3061 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3121 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3181 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3241 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3301 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3361 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3421 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3481 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3541 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3601 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3661 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3721 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3781 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3841 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3901 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3961 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4021 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4081 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4141 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4201 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4261 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4321 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4381 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4441 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4561 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4621 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4681 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4741 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4801 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4861 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4921 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4981 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5041 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5101 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5161 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5221 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5281 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5341 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5401 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5461 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5521 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5581 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5641 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5701 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5761 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5821 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5881 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5941 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6001 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6061 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6121 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6181 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6241 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6301 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6361 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6421 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6481 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6541 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6601 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6661 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6721 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6781 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6841 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8041 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8101 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8161 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8221 gacaattgca tgaagaatct gcttagg
//
