LOCUS       V026515                 8310 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026515
VERSION     V026515
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..2990
                     /label="RPS6KA6(NM_014496)"
                     /note="RPS6KA6(NM_014496)"
                     /gene="RPS6KA6"
     CDS             2997..3062
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3086..3674
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3740..3788
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3990..4418
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4432..4761
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4828..5622
                     /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    5798..5919
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5968..5984)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5992..6008
                     /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(6016..6046)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6061..6082
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6370..6958)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7129..7989)
                     /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(7990..8094)
                     /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 ccaccatgct accattcgct cctcaggacg
      781 agccctggga ccgagaaatg gaagtgttca gcggcggcgg cgcgagcagc ggcgaggtaa
      841 atggtcttaa aatggttgat gagccaatgg aagagggaga agcagattct tgtcatgatg
      901 aaggagttgt taaagaaatc cctattactc atcatgttaa ggaaggctat gagaaagcag
      961 atcctgcaca gtttgagttg ctcaaggttc ttggtcaggg gtcatttgga aaggtttttc
     1021 ttgttagaaa gaagaccggt cctgatgctg ggcagctcta tgcaatgaag gtgttaaaaa
     1081 aagcctcttt aaaagttcga gacagagttc ggacaaagat ggagagggat atactggtgg
     1141 aagtaaatca tccatttatt gtcaaattgc actatgcctt tcagactgaa gggaaactgt
     1201 acttaatact ggattttctc aggggaggag atgttttcac aagattatcc aaagaggttc
     1261 tgtttacaga ggaagatgtg aaattctacc tcgcagaact ggcccttgct ttggatcatc
     1321 tgcaccaatt aggaattgtt tatagagacc tgaagccaga aaacattttg cttgatgaaa
     1381 taggacatat caaattaaca gattttggac tcagcaagga gtcagtagat caagaaaaga
     1441 aggcttactc attttgtggt acagtagagt atatggctcc tgaagtagta aataggagag
     1501 gccattccca gagtgctgat tggtggtcat atggtgttct tatgtttgaa atgcttactg
     1561 gtactctgcc atttcaaggt aaagacagaa atgagaccat gaatatgata ttaaaagcaa
     1621 aacttggaat gcctcaattt cttagtgctg aagcacaaag tcttctaagg atgttattca
     1681 aaaggaatcc agcaaataga ttgggatcag aaggagttga agaaatcaaa agacatctgt
     1741 tttttgcaaa tattgactgg gataaattat ataaaagaga agttcaacct cctttcaaac
     1801 ctgcttctgg aaaaccagat gatacttttt gttttgatcc tgaatttact gcaaaaacac
     1861 ctaaagattc tcccggtttg ccagccagtg caaatgctca tcagctcttc aaaggattca
     1921 gctttgttgc aacttctatt gcagaagaat ataaaatcac tcctatcaca agtgcaaatg
     1981 tattaccaat tgttcagata aatggaaatg ctgcacaatt tggtgaagta tatgaattga
     2041 aggaggatat tggtgttggc tcctactctg tttgcaagcg atgcatacat gcaactacca
     2101 acatggaatt tgcagtgaag atcattgaca aaagtaagcg agacccttca gaagagattg
     2161 aaatattgat gcgctatgga caacatccca acattattac tttgaaggat gtctttgatg
     2221 atggtagata tgtttacctt gttacggatt taatgaaagg aggagagtta cttgaccgta
     2281 ttctcaaaca aaaatgtttc tcggaacggg aggctagtga tatactatat gtaataagta
     2341 agacagttga ctatcttcat tgtcaaggag ttgttcatcg tgatcttaaa cctagtaata
     2401 ttttatacat ggatgaatca gccagtgcag attcaatcag gatatgtgat tttgggtttg
     2461 caaaacaact tcgaggagaa aatggacttc tcttaactcc atgctacact gcaaactttg
     2521 ttgcacctga ggttcttatg caacagggat atgatgctgc ttgtgatatc tggagtttag
     2581 gagtcctttt ttacacaatg ttggctggct acactccatt tgctaatggc cccaatgata
     2641 ctcctgaaga gatactgctg cgtataggca atggaaaatt ctctttgagt ggtggaaact
     2701 gggacaatat ttcagacgga gcaaaggatt tgctttccca tatgcttcat atggacccac
     2761 atcagcggta tactgctgaa caaatattaa agcactcatg gataactcac agagaccagt
     2821 tgccaaatga tcagccaaag agaaatgatg tgtcacatgt tgttaaggga gcaatggttg
     2881 caacatactc tgccctgact cacaagacct ttcaaccagt cctagagcct gtagctgctt
     2941 caagcttagc ccagcgacgg agcatgaaaa agcgaacatc aactggcctg ggatccgact
     3001 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     3061 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3121 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3181 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3241 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3301 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3361 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3421 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3481 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3541 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3601 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3661 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     3721 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     3781 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3841 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3901 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     3961 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4021 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4081 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4141 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4201 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4261 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4321 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     4381 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4441 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4501 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4561 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4621 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4681 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4741 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4801 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4861 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4921 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4981 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5041 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5101 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5161 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5221 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5281 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5341 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5401 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5461 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5521 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5581 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5641 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5701 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5761 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5881 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5941 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6001 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6061 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6121 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6181 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6241 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6301 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6541 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     6601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6781 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6841 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6901 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8101 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     8161 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     8221 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     8281 accgacaatt gcatgaagaa tctgcttagg
//
