LOCUS       V018686                 9685 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018686
VERSION     V018686
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             651..5489
                     /label="LRP6(NM_002336)"
                     /note="LRP6(NM_002336)"
                     /gene="LRP6"
     CDS             5496..5513
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
     CDS             5526..5609
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     CDS             5631..6350
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    6473..6594
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(6601..7056)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7083..7187
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        7189..7546
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7581..8375
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    8607..8654
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      8983..9571
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacgccacc atgggggccg
      661 tcctgaggag cctcctggcc tgcagcttct gtgtgctcct gagagcggcc cctttgttgc
      721 tttatgcaaa cagacgggac ttgcgattgg ttgatgctac aaatggcaaa gagaatgcta
      781 cgattgtagt tggaggcttg gaggatgcag ctgcggtgga ctttgtgttt agtcatggct
      841 tgatatactg gagtgatgtc agcgaagaag ccattaaacg aacagaattt aacaaaactg
      901 agagtgtgca gaatgttgtt gtttctggat tattgtcccc cgatgggctg gcatgtgatt
      961 ggcttggaga aaaattgtac tggacagatt ctgaaactaa tcggattgaa gtttctaatt
     1021 tagatggatc tttacgaaaa gttttatttt ggcaagagtt ggatcaaccc agagctattg
     1081 ccttagatcc ttcaagtggg ttcatgtact ggacagactg gggagaagtg ccaaagatag
     1141 aacgtgctgg aatggatggt tcaagtcgct tcattataat aaacagtgaa atttactggc
     1201 caaatggact gactttggat tatgaagaac aaaagcttta ttgggcagat gcaaaactta
     1261 atttcatcca caaatcaaat ctggatggaa caaatcggca ggcagtggtt aaaggttccc
     1321 ttccacatcc ttttgccttg acgttatttg aggacatatt gtactggact gactggagca
     1381 cacactccat tttggcttgc aacaagtata ctggtgaggg tctgcgtgaa atccattctg
     1441 acatcttctc tcccatggat atacatgcct tcagccaaca gaggcagcca aatgccacaa
     1501 atccatgtgg aattgacaat gggggttgtt cccatttgtg tttgatgtct ccagtcaagc
     1561 ctttttatca gtgtgcttgc cccactgggg tcaaactcct ggagaatgga aaaacctgca
     1621 aagatggtgc cacagaatta ttgcttttag ctcgaaggac agacttgaga cgcatttctt
     1681 tggatacacc agattttaca gacattgttc tgcagttaga agacatccgt catgccattg
     1741 ccatagatta cgatcctgtg gaaggctaca tctactggac tgatgatgaa gtgagggcca
     1801 tacgccgttc atttatagat ggatctggca gtcagtttgt ggtcactgct caaattgccc
     1861 atcctgatgg tattgctgtg gactgggttg cacgaaatct ttattggaca gacactggca
     1921 ctgatcgaat agaagtgaca aggctcaatg ggaccatgag gaagatcttg atttcagagg
     1981 acttagagga accccgggct attgtgttag atcccatggt tgggtacatg tattggactg
     2041 actggggaga aattccgaaa attgagcgag cagctctgga tggttctgac cgtgtagtat
     2101 tggttaacac ttctcttggt tggccaaatg gtttagcctt ggattatgat gaaggcaaaa
     2161 tatactgggg agatgccaaa acagacaaga ttgaggttat gaatactgat ggcactggga
     2221 gacgagtact agtggaagac aaaattcctc acatatttgg atttactttg ttgggtgact
     2281 atgtttactg gactgactgg cagaggcgta gcattgaaag agttcataaa cgaagtgcag
     2341 agagggaagt gatcatagat cagctgcctg acctcatggg cctaaaggct acaaatgttc
     2401 atcgagtgat tggttccaac ccctgtgctg aggaaaacgg gggatgtagc catctctgcc
     2461 tctatagacc tcagggcctt cgctgtgctt gccctattgg ctttgaactc atcagtgaca
     2521 tgaagacctg cattgtccca gaggctttcc ttttgttttc acggagagca gatatcagac
     2581 gaatttctct ggaaacaaac aataataatg tggctattcc actcactggt gtcaaagaag
     2641 cttctgcttt ggattttgat gtgacagaca accgaattta ttggactgat atatcactca
     2701 agaccatcag cagagccttt atgaatggca gtgcactgga acatgtggta gaattcggct
     2761 tagattatcc agaaggcatg gcagtagact ggcttgggaa gaacttgtac tgggcagaca
     2821 caggaacgaa tcgaattgag gtgtcaaagt tggatgggca gcaccgacaa gttttggtgt
     2881 ggaaagacct agatagtccc agagctctcg cgttggaccc tgccgaagga tttatgtatt
     2941 ggactgaatg gggtggaaaa cctaagatag acagagctgc aatggatgga agtgaacgta
     3001 ctaccttagt tccaaatgtg gggcgggcaa acggcctaac tattgattat gctaaaagga
     3061 ggctttattg gacagacctg gacaccaact taatagaatc ttcaaatatg cttgggctca
     3121 accgtgaagt tatagcagat gacttgcctc atccttttgg cttaactcag taccaagatt
     3181 atatctactg gacggactgg agccgacgca gcattgagcg tgccaacaaa accagtggcc
     3241 aaaaccgcac catcattcag ggccatttgg attatgtgat ggacatcctc gtctttcact
     3301 catctcgaca gtcagggtgg aatgaatgtg cttccagcaa tgggcactgc tcccacctct
     3361 gcttggctgt gccagttggg ggttttgttt gtggatgccc tgcccactac tctcttaatg
     3421 ctgacaacag gacttgtagt gctcctacga ctttcctgct cttcagtcaa aagagtgcca
     3481 tcaaccgcat ggtgattgat gaacaacaga gccccgacat catccttccc atccacagcc
     3541 ttcggaatgt ccgggccatt gactatgacc cactggacaa gcaactctat tggattgact
     3601 cacgacaaaa catgatccga aaggcacaag aagatggcag ccagggcttt actgtggttg
     3661 tgagctcagt tccgagtcag aacctggaaa tacaacccta tgacctcagc attgatattt
     3721 acagccgcta catctactgg acttgtgagg ctaccaatgt cattaatgtg acaagattag
     3781 atgggagatc agttggagtg gtgctgaaag gcgagcagga cagacctcga gccgttgtgg
     3841 taaacccaga gaaagggtat atgtatttta ccaatcttca ggaaaggtct cctaaaattg
     3901 aacgggctgc tttggatggg acagaacggg aggtcctctt tttcagtggc ttaagtaaac
     3961 caattgcttt agcccttgat agcaggctgg gcaagctctt ttgggctgat tcagatctcc
     4021 ggcgaattga aagcagtgat ctctcaggtg ctaaccggat agtattagaa gactccaata
     4081 tcttgcagcc tgtgggactt actgtgtttg aaaactggct ctattggatt gataaacagc
     4141 agcaaatgat tgaaaaaatt gacatgacag gtcgagaggg tagaaccaaa gtccaagctc
     4201 gaattgccca gcttagtgac attcatgcag taaaggagct gaaccttcaa gaatacagac
     4261 agcacccttg tgctcaggat aatggtggct gttcacatat ttgtcttgta aagggggatg
     4321 gtactacaag gtgttcttgc cccatgcacc tggttctact tcaagatgag ctatcatgtg
     4381 gagaacctcc aacatgttct cctcagcagt ttacttgttt cacgggggaa attgactgta
     4441 tccctgtggc ttggcggtgc gatgggttta ctgaatgtga agaccacagt gatgaactca
     4501 attgtcctgt atgctcagag tcccagttcc agtgtgccag tgggcagtgt attgatggtg
     4561 ccctccgatg caatggagat gcaaactgcc aggacaaatc agatgagaag aactgtgaag
     4621 tgctttgttt aattgatcag ttccgctgtg ccaatggtca gtgcattgga aagcacaaga
     4681 agtgtgatca taatgtggat tgcagtgaca agtcagatga actggattgt tatccgactg
     4741 aagaaccagc accacaggcc accaatacag ttggttctgt tattggcgta attgtcacca
     4801 tttttgtgtc tggaactgta tactttatct gccagaggat gttgtgtcca cgtatgaagg
     4861 gagatgggga aactatgact aatgactatg tagttcatgg accagcttct gtgcctcttg
     4921 gttatgtgcc acacccaagt tctttgtcag gatctcttcc aggaatgtct cgaggtaaat
     4981 caatgatcag ctccctcagt atcatggggg gaagcagtgg acccccctat gaccgagccc
     5041 atgttacagg agcatcatca agtagttctt caagcaccaa aggcacttac ttccctgcaa
     5101 ttttgaaccc tccaccatcc ccagccacag agcgatcaca ttacactatg gaatttggat
     5161 attcttcaaa cagtccttcc actcataggt catacagcta caggccatat agctaccggc
     5221 actttgcacc ccccaccaca ccctgcagca cagatgtttg tgacagtgac tatgctccta
     5281 gtcggagaat gacctcagtg gcaacagcca agggctatac cagtgacttg aactatgatt
     5341 cagaacctgt gcccccacct cccacacccc gaagccaata cttgtcagca gaggagaact
     5401 atgaaagctg cccaccttct ccatacacag agaggagcta ttctcatcac ctctacccac
     5461 cgccaccctc tccctgtaca gactcctccg gtaccctggt gccgcgcggc agccatatgg
     5521 ctagctggag ccatccgcag tttgaaaaag gtggtggtag cggtggtggt tcaggtggta
     5581 gtgcatggtc acaccctcag tttgagaaac tggatccacc ggtcgccacc atggtgagca
     5641 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     5701 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     5761 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     5821 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     5881 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     5941 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     6001 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     6061 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     6121 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     6181 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     6241 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     6301 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     6361 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     6421 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     6481 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6541 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     6601 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     6661 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     6721 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     6781 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     6841 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     6901 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     6961 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     7021 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     7081 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7141 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     7201 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     7261 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     7321 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     7381 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     7441 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     7501 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     7561 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     7621 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     7681 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     7741 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     7801 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     7861 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     7921 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     7981 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     8041 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     8101 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     8161 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     8221 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     8281 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     8341 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     8401 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     8461 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     8521 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     8581 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     8641 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     8701 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     8761 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     8821 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     8881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     8941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     9001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     9061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     9121 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     9181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     9241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     9301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     9361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     9421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     9481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     9541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     9601 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     9661 ggataaccgt attaccgcca tgcat
//
