LOCUS       V046727                 9219 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046727
VERSION     V046727
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..3899
                     /label="PLEKHA6(NM_014935)"
                     /note="PLEKHA6(NM_014935)"
                     /gene="PLEKHA6"
     CDS             3906..3971
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3995..4583
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4649..4697
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4899..5327
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5341..5670
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5737..6531
                     /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    6707..6828
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6877..6893)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6901..6917
                     /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(6925..6955)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6970..6991
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7279..7867)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8038..8898)
                     /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(8899..9003)
                     /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 ccaccatgtc caataaaaca ggtgggaaac
      781 gcccggctac caccaacagt gacataccca accacaacat ggtgtccgag gtccctccag
      841 agcggcccag cgtccgggca actcgcacag cccgcaaagc cgtcgccttt ggcaagcgct
      901 cacactccat gaagcggaac cccaatgcac ctgtcaccaa ggcgggctgg ctcttcaaac
      961 aggccagctc cggggttaag cagtggaaca agcgctggtt cgtcctggtg gatcgctgcc
     1021 tcttctacta taaagatgag aaggaagaga gtatcctggg cagcatcccc ctcctgagct
     1081 tccgggtagc cgcagtgcag ccctcagaca acatcagccg gaaacacacg tttaaggctg
     1141 agcatgccgg ggtccgcacc tacttcttca gtgccgagag ccccgaggag caagaggcct
     1201 ggatccaggc catgggggag gctgctcgag tacagatccc tccagcccag aagtcagtgc
     1261 cccaagctgt gcggcacagc catgagaagc cagactcgga gaacgtccca cccagcaagc
     1321 accaccagca gccaccccac aacagcctcc ctaagcctga gccagaggcc aagactcgag
     1381 gggagggtga tggccgaggc tgtgagaagg cagagagaag gcctgagagg ccagaagtca
     1441 agaaagagcc tccggtgaaa gccaatggcc tcccagctgg accggagcca gcctcagagc
     1501 cgggcagccc ttaccccgag ggcccaagag tgccaggggg tggggaacag cctgcccagc
     1561 ccaatggctg gcagtaccac tccccaagcc ggccagggag cacagctttc ccgtctcagg
     1621 atggagagac tgggggacac cggcggagtt tcccaccacg caccaaccct gacaaaattg
     1681 cccagcgcaa gagctccatg aaccagcttc agcagtgggt gaatctgcgc cggggggtac
     1741 ccccgcctga agaccttcgg agtccctcta ggttctatcc tgtgtctcgc agggtccctg
     1801 agtactatgg cccctactcc tcccagtacc ccgatgatta tcagtactac ccgccaggag
     1861 tgcggccgga gagcatctgt tccatgccgg cctatgatcg gatcagcccg ccctgggccc
     1921 tggaggacaa gcgccatgcc ttccgcaatg ggggtggccc tgcctaccag ctgcgagagt
     1981 ggaaggagcc cgccagctac gggcggcagg atgccaccgt ctggatccca agcccctccc
     2041 ggcagccagt ctattatgat gagctggatg ccgcctctag ctccctgcgc cgcctgtccc
     2101 tgcagccccg ctcccactct gtgccccgct cacccagcca gggctcctac agccgtgccc
     2161 gcatttactc ccctgtccgc tcacccagtg cccgttttga gcggctgcca cctcgcagtg
     2221 aggacatcta tgctgaccct gctgcctatg tgatgaggcg atccatcagc tcccccaagg
     2281 tccctccata cccagaagtg ttccgggaca gcctccacac ctacaagtta aacgagcaag
     2341 acacagataa gctgctggga aaattgtgtg agcagaacaa ggtggtgagg gagcaggacc
     2401 ggctggtgca gcagctccga gctgagaagg agagcctgga aagtgccttg atggggaccc
     2461 accaggagct ggagatgttt ggaagccagc ccgcctaccc agaaaagctg cgacacaaaa
     2521 aggattcact gcagaaccag ctcatcaaca tccgcgtgga gctgtctcag gcgaccacgg
     2581 ccctgacaaa cagcaccata gagtatgagc acctcgagtc tgaggtctct gccctgcacg
     2641 atgacctctg ggagcagctc aatttggaca cccagaatga ggtgctgaac cggcaaatcc
     2701 aaaaggagat ctggaggatc caggacgtga tggaggggct gaggaagaac aacccctccc
     2761 ggggcacgga caccgccaag cacagaggag gacttggccc ctcagccacc tacagctcca
     2821 acagcccggc cagccccctc agctctgcca gcctcaccag ccccctgagc cccttttcac
     2881 tggtgtcggg ctctcagggg tcccccacca agcctggctc caacgagccc aaggcaaact
     2941 atgaacaaag caagaaagac ccccaccaga cattgcccct ggacaccccc agagacatca
     3001 gccttgtgcc caccaggcaa gaggtagagg cagagaagca ggcagctctc aacaaagttg
     3061 gcgttgtgcc ccctcggaca aaatcgccca ctgatgatga ggtgacccca tcagcagtgg
     3121 taagaaggaa tgccagtggg ctcaccaatg gactctcctc ccaggaacgc cccaagagtg
     3181 ctgtgtttcc tggcgagggg aaggtcaaga tgagcgtgga ggagcagatt gaccgaatgc
     3241 ggcggcacca gagtggctcc atgagggaga agcggaggag cctgcagctc ccggccagcc
     3301 cggcccccga ccccagtccc cggccagcct acaaagtggt gcgccgccac cgcagcatcc
     3361 atgaggtaga catctccaac ctggaggcag ccctgcgggc agaggagcct ggcgggcatg
     3421 cctacgagac accccgggag gaaattgccc ggcttcgcaa aatggagcta gagccccagc
     3481 attatgacgt ggacatcaat aaggagctct ccactccaga caaagtcctc atccctgaac
     3541 ggtacattga cctggagcct gacactcccc tgagccctga ggagttgaag gagaagcaga
     3601 agaaggtgga gaggatcaag acactcattg ccaaatccag tatgcagaac gtggtgccca
     3661 tcggcgaggg ggactctgtg gacgtgcccc aggactcaga gagccagctg caggagcagg
     3721 agaagcggat tgaaatctcc tgcgccctgg cgaccgaggc ctcccgcagg ggccgcatgc
     3781 tgtctgtgca atgtgccacc ccaagccctc ccacctcccc tgcttccccg gctcctccag
     3841 caaaccccct gtcgtctgaa tccccacggg gcgccgacag cagctatacc atgcgggtcg
     3901 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3961 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     4021 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4081 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4141 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4201 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4261 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4321 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4381 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4441 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4501 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4561 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     4621 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     4681 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4741 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4801 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4861 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4921 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4981 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5041 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5101 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5161 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5221 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     5281 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5341 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5401 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5461 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5521 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5581 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5641 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5701 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5761 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5821 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5881 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5941 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6001 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6061 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6121 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6181 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6241 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6301 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6361 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6421 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6481 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6541 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6601 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6661 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6721 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6781 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6841 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6901 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6961 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7021 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7081 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7441 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7681 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8101 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8161 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8221 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8281 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8341 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8401 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8461 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8521 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8581 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8641 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8701 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8761 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8821 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8881 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8941 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9001 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9061 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9121 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9181 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
