LOCUS       V040275                 9891 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040275
VERSION     V040275
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..4571
                     /label="PLEKHA7(NM_001410960)"
                     /note="PLEKHA7(NM_001410960)"
                     /gene="PLEKHA7"
     CDS             4578..4643
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4667..5255
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5321..5369
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5571..5999
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6013..6342
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6409..7203
                     /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    7379..7500
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7549..7565)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7573..7589
                     /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(7597..7627)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7642..7663
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7951..8539)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8710..9570)
                     /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(9571..9675)
                     /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 ggcggcgacg gtcgggcggg
      781 acactttacc tgagcattgg tcctacgggg tgtgccggga tggccgcgtc ttcttcatca
      841 atgaccagct ccgctgcacg acctggctgc atccgcgcac cggggagccc gtcaactcgg
      901 gccacatgat ccgctcagac ctgccccgcg gctgggagga gggcttcacg gaggagggcg
      961 ccagctactt catcgaccat aaccagcaga ccacagcatt caggcatcct gtgacgggac
     1021 agttttctcc agaaaatagt gaattcattc ttcaagaaga gccgaatcca catatgtcga
     1081 agcaagacag aaaccaaaga ccgtccagca tggtcagtga aacatccacg gctgggaccg
     1141 cctccaccct ggaggccaag cctggaccca agatcataaa gtccagcagt aaagtccaca
     1201 gctttgggaa gagagaccag gccattcgga ggaaccccaa tgttcccgtg gtggtgaggg
     1261 gctggctgca caagcaggac agttctggga tgaggctgtg gaaaaggagg tggtttgtgc
     1321 ttgctgatta ctgcttattt tactataaag acagccgaga agaagcggtc ctcgggagca
     1381 tccccttgcc cagctacgtg atctctcctg tggcccctga ggatcgcata agccgcaaat
     1441 attcctttaa ggctgtgcac acggggatgc gagcgctcat ctataacagc tccacagcgg
     1501 gctctcaggc cgagcagtca ggcatgagga cctactactt cagtgccgac acccaggagg
     1561 acatgaacgc ttgggtcagg gccatgaacc aggctgcaca ggtgctgtct cgatcgtcac
     1621 tgaagaggga tatggagaag gtggagcggc aggctgtccc ccaggccaac cacacagagt
     1681 cctgtcacga atgtggccgg gtgggacccg gacatacgag agattgtcct catcgtggcc
     1741 atgatgacat tgtcaacttc gagaggcagg agcaggaggg agagcagtac cgttcccaga
     1801 gggacccact ggagggcaag cgggaccgga gcaaggccag gtctccgtac tcgccagccg
     1861 aggaggatgc cttgtttatg gatttaccca ctggcccaag aggccagcag gcacagcccc
     1921 aacgggcaga gaagaatgga atgctgcctg cctcatatgg cccaggagaa cagaatggga
     1981 ctggtgggta ccagcgggcc tttcctccca ggaccaaccc tgaaaaacac agccaaagga
     2041 agagcaatct ggcccaggtg gagcactggg caagggccca gaaaggggat agcaggagtc
     2101 ttcccttgga ccagacgctt cctcgccagg gtcctggcca atccctgtcc ttcccagaaa
     2161 actaccagac tcttcccaag agcacccgac acccctcggg gggctcctcg ccacctcccc
     2221 gaaacctgcc aagtgactac aagtatgcgc aggaccgagc cagccacctg aagatgtcga
     2281 gtgaagagcg ccgggcgcac cgggatggca ccgtgtggca gctctacgag tggcagcagc
     2341 gccagcagtt ccggcacggc agccccacag cgcccatctg ccttggctcc ccagagttca
     2401 ccgaccaggg ccggagcagg agcatgctag aggtgccccg ctccatctct gtgcctccat
     2461 ctccctcgga catccctccc ccaggacccc caagggtctt cccaccccgg cggccacaca
     2521 caccagcaga gcgagtcaca gtgaagccac cggaccagag gaggagtgtg gacatctcgc
     2581 tgggggattc tccaaggagg gcacggggcc acgctgtcaa gaactcatct catgtggacc
     2641 gacgctccat gccctccatg ggttacatga cccacaccgt cagcgctccc agtttacatg
     2701 gaaaatcggc tgatgatacc tacctccagc tgaagaaaga cctggagtac ctggatctaa
     2761 agatgacagg ccgggacctt ctcaaggatc gaagtctgaa gcctgtgaag atcgctgaga
     2821 gcgacactga cgtcaaactg agcatcttct gtgaacaaga cagggtcctc caggacttgg
     2881 aagacaagat acgagccctt aaagagaaca aagaccagct agaatctgtg ctggaggtgt
     2941 tgcacagaca gatggagcag taccgagacc agccccagca cttggagaag attgcctacc
     3001 agcagaagtt gctgcaggag gaccttgtcc atatccgagc tgagctctcc agagagtcca
     3061 ctgagatgga aaatgcttgg aacgaatacc tgaagttgga gaatgatgtg gaacagctga
     3121 agcagaccct gcaggagcaa cacagaagag cctttttttt ccaggagaaa tcgcagatac
     3181 agaaagatct atggagaatt gaagatgtca ctgcaggcct gagtgcaaat aaagagaact
     3241 tcagaattct agtggagtca gtaaaaaatc cggagagaaa aacggtgcct ttgtttcctc
     3301 acccgcctgt gccttcactc tcaacttctg agagcaagcc gcccccacag cccagtcctc
     3361 ccaccagccc tgtgcggacc cctctggagg ttcgactctt cccccagctg caaacctacg
     3421 tgccgtaccg acctcaccca ccccagctga ggaaagtgac atcccccctt cagtcaccaa
     3481 ctaaggcgaa gcccaaagtt caggaagatg aagcacctcc caggccccca ctccccgaac
     3541 tctacagccc agaggaccag cccccggctg tgccgcctct gccaagagag gccaccatca
     3601 tccggcacac atctgtgcgg ggcctcaagc ggcagtcaga cgagaggaag cgagaccggg
     3661 agctggggca gtgtgtgaat ggggattcca gggtggagct gcggtcgtat gtcagtgagc
     3721 ctgagctggc gaccctcagc ggggacatgg cccagccctc cctaggactt gtgggccctg
     3781 agagcaggta ccagacgctg ccaggcagag ggctctcagg gtccacgtca aggctccagc
     3841 agtcgtccac cattgctccc tacgtcacac tccggagggg tctcaatgcc gaaagcagca
     3901 aggcgacctt ccctagacct aagagtgcct tggagcgcct gtactcaggg gatcaccagc
     3961 gaggcaagat gagtgcagag gagcagctgg agcgcatgaa gcgacaccag aaggccctgg
     4021 tccgagagcg caagaggaca ctgggccaag gggagaggac gggcctgccc tcatctcgct
     4081 acctcagccg gccgctccct ggagatcttg gctcatggaa gcgtgagcag gactttgacc
     4141 tgcagttgct ggaacgggtc gtgcaagggg aaaaaaagga caaggaggag aatggctggt
     4201 tgaaagtgca ggccatgcct gtcactgagt tggacctgga gcctcaagac tatgacttgg
     4261 acatcagcag agagctgtcc aaaccagaga aggtgtcaat ccctgagcgc tacgtggagc
     4321 tagatcccga agagccaccc agccttgagg agctgcaggc gcggtaccgc aaagccgaga
     4381 agatccgcaa catcctcgcc cggtcaagca tgtgcaacct acagccgacc tcaggccagg
     4441 accagaacag tgtggctgac ctggacttgc agctgcagga gcaggagcgc atcatcaaca
     4501 tctcctacgc cctggcctcc gaggcctccc agcgtagcaa gcaggtggca gcccaggcag
     4561 tgactgaccc gggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     4621 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     4681 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4741 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4801 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4861 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4921 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4981 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5041 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5101 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5161 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5221 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     5281 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5341 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5401 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5461 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5521 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     5581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5701 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     5761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5941 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     6001 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     6061 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     6121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     6181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6241 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     6301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     6361 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     6421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     7021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     7081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     7141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     7201 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     7261 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     7321 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     7381 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7441 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7501 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     7561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     8041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     8101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     8161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     8221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     8281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     8341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     8401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8461 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     9001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     9061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     9121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     9181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9721 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     9781 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     9841 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
