LOCUS       V034325                 8603 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034325
VERSION     V034325
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             625..4497
                     /label="PLCG1(NM_002660)"
                     /note="PLCG1(NM_002660)"
                     /gene="PLCG1"
     CDS             4549..5268
                     /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    5391..5512
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5519..5974)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6001..6105
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        6107..6464
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6499..7293
                     /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    7525..7572
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      7901..8489
                     /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 atctcgaggc caccatggcg ggcgccgcgt ccccttgcgc caacggctgc
      661 gggcccggcg cgccctcgga cgccgaggtg ctgcacctct gccgcagcct cgaggtgggc
      721 accgtcatga ctttgttcta ctccaagaag tcgcagcgac ccgagcggaa gaccttccag
      781 gtcaagctgg agacgcgcca gatcacgtgg agccggggcg ccgacaagat cgagggggcc
      841 attgacattc gtgaaattaa ggagatccgc ccagggaaga cctcacggga ctttgatcgc
      901 tatcaagagg acccagcttt ccggccggac cagtcacatt gctttgtcat tctctatgga
      961 atggaatttc gcctgaaaac gctgagcctg caagccacat ctgaggatga agtgaacatg
     1021 tggatcaagg gcttaacttg gctgatggag gatacattgc aggcacccac acccctgcag
     1081 attgagaggt ggctccggaa gcagttttac tcagtggatc ggaatcgtga ggatcgtata
     1141 tcagccaagg acctgaagaa catgctgtcc caggtcaact accgggtccc caacatgcgc
     1201 ttcctccgag agcggctgac ggacctggag cagcgcagcg gggacatcac ctacgggcag
     1261 tttgctcagc tgtaccgcag cctcatgtac agcgcccaga agacgatgga cctccccttc
     1321 ttggaagcca gtactctgag ggctggggag cggccggagc tttgccgagt gtcccttcct
     1381 gagttccagc agttccttct tgactaccag ggggagctgt gggctgttga tcgcctccag
     1441 gtgcaggagt tcatgctcag cttcctccga gaccccttac gagagatcga ggagccatac
     1501 ttcttcctgg atgagtttgt caccttcctg ttctccaaag agaacagtgt gtggaactcg
     1561 cagctggatg cagtatgccc ggacaccatg aacaaccctc tttcccacta ctggatctcc
     1621 tcctcgcaca acacgtacct gaccggggac cagttctcca gtgagtcctc cttggaagcc
     1681 tatgctcgct gcctgcggat gggctgtcgc tgcattgagt tggactgctg ggacggcccg
     1741 gatgggatgc cagttattta ccatgggcac acccttacca ccaagatcaa gttctcagat
     1801 gtcctgcaca ccatcaagga gcatgccttt gtggcctcag agtacccagt catcctgtcc
     1861 attgaggacc actgcagcat tgcccagcag agaaacatgg cccaatactt caagaaggtg
     1921 ctgggggaca cactcctcac caagcccgtg gagatctctg ccgacgggct cccctcaccc
     1981 aaccagctta agaggaagat cctcatcaag cacaagaagc tggctgaggg cagtgcctac
     2041 gaggaggtgc ctacatccat gatgtactct gagaacgaca tcagcaactc tatcaagaat
     2101 ggcatcctct acctggagga ccctgtgaac cacgaatggt atccccacta ctttgttctg
     2161 accagcagca agatctacta ctctgaggag accagcagtg accagggcaa cgaggatgag
     2221 gaggagccca aggaggtcag cagcagcaca gagctgcact ccaatgagaa gtggttccat
     2281 gggaagctag gggcagggcg tgacgggcgt cacatcgctg agcgcctgct tactgagtac
     2341 tgcatcgaga ccggagcccc tgacggctcc ttcctcgtgc gagagagtga gaccttcgtg
     2401 ggcgactaca cgctctcttt ctggcggaac gggaaagtcc agcactgccg tatccactcc
     2461 cggcaagatg ctgggacccc caagttcttc ttgacagaca acctcgtctt tgactccctc
     2521 tatgacctca tcacgcacta ccagcaggtg cccctgcgct gtaatgagtt tgagatgcga
     2581 ctttcagagc ctgtcccaca gaccaacgcc cacgagagca aagagtggta ccacgcgagc
     2641 ctgaccagag cacaggctga gcacatgcta atgcgcgtcc ctcgtgatgg ggccttcctg
     2701 gtgcggaagc ggaatgaacc caactcatat gccatctctt tccgggctga gggcaagatc
     2761 aagcattgcc gtgtccagca agagggccag acagtgatgc tagggaactc ggagttcgac
     2821 agccttgttg acctcatcag ctactatgag aaacacccgc tataccgcaa gatgaagctg
     2881 cgctatccca tcaacgagga ggcactggag aagattggca cagctgagcc tgactacggg
     2941 gccctgtatg agggacgcaa ccctggcttc tatgtagagg caaaccctat gccaactttc
     3001 aagtgtgcag tcaaagccct ctttgactac aaggcccaga gggaggacga gctgaccttc
     3061 atcaagagcg ccatcatcca gaatgtggag aagcaagagg gaggctggtg gcgaggggac
     3121 tacggaggga agaagcagct gtggttccca tcaaactacg tggaagagat ggtcaacccc
     3181 gtggccctgg agccggagag ggagcacttg gacgagaaca gccccctagg ggacttgctg
     3241 cggggggtct tggatgtgcc ggcttgtcag attgccatcc gtcctgaggg caagaacaac
     3301 cggctcttcg tcttctccat cagcatggcg tcggtggccc actggtccct ggatgttgct
     3361 gccgactcac aggaggagct gcaggactgg gtgaaaaaga tccgtgaagt ggcccagaca
     3421 gcagacgcca ggctcactga agggaagata atggaacgga ggaagaagat tgccctggag
     3481 ctctctgaac ttgtcgtcta ctgccggcct gttccctttg atgaagagaa gattggcaca
     3541 gaacgtgctt gctaccggga catgtcatcc ttcccggaaa ccaaggctga gaaatacgtg
     3601 aacaaggcca aaggcaagaa gttccttcag tacaatcgac tgcagctctc ccgcatctac
     3661 cccaagggcc agcgactgga ttcctccaac tacgatcctt tgcccatgtg gatctgtggc
     3721 agtcagcttg tggccctcaa cttccagacc cctgacaagc ctatgcagat gaaccaggcc
     3781 ctcttcatga cgggcaggca ctgtggctac gtgctgcagc caagcaccat gcgggatgag
     3841 gccttcgacc cctttgacaa gagcagcctc cgcgggctgg agccatgtgc catctctatt
     3901 gaggtgctgg gggcccgaca tctgccaaag aatggccgag gcattgtgtg tccttttgtg
     3961 gagattgagg tggctggagc tgagtatgac agcaccaagc agaagacaga gtttgtggtg
     4021 gacaatggac tcaaccctgt atggccagcc aagcccttcc acttccagat cagtaaccct
     4081 gaatttgcct ttctgcgctt cgtggtgtat gaggaagaca tgtttagtga ccagaatttc
     4141 ctggctcagg ctactttccc agtaaaaggc ctgaagacag gatacagagc agtgcctttg
     4201 aagaacaact acagtgagga cctggagttg gcctccctgc tgatcaagat tgacattttc
     4261 cctgccaagc aggagaatgg tgacctcagt cccttcagtg gtacgtccct gcgggagcgg
     4321 ggctcagatg cctcaggcca gctgtttcat ggccgagccc gggaaggctc ctttgaatcc
     4381 cgctaccagc agccgtttga ggacttccgc atctcccagg agcatctcgc agaccatttt
     4441 gacagtcgag aacgaagggc cccaagaagg actcgggtca atggagacaa ccgcctcggc
     4501 ggcggaggga gtggaggagg gggaagcttg gatccaccgg tcgccaccat ggtgagcaag
     4561 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4621 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     4681 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4741 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     4801 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     4861 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4921 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4981 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     5041 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     5101 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     5161 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     5221 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac
     5281 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     5341 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     5401 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5461 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     5521 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5581 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5641 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5701 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5761 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5821 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5881 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5941 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     6001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     6121 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     6181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     6241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     6301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     6481 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6541 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6601 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6661 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     6721 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6781 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6841 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6901 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6961 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     7021 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     7081 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7141 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7201 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7261 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7321 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7381 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7441 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     7501 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     7561 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     7621 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     7681 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     7741 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7801 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     8041 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     8341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     8401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     8461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8521 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     8581 ataaccgtat taccgccatg cat
//
