LOCUS       V038776                 6990 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038776
VERSION     V038776
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..2241
                     /label="Ripk2(NM_138952)"
                     /note="Ripk2(NM_138952)"
                     /gene="Ripk2"
     CDS             2242..2307
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2349..2935
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             2936..3655
                     /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    3778..3899
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3906..4361)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4388..4492
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4494..4851
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4886..5680
                     /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    5912..5959
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6288..6876
                     /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 caccatgaac ggggacgcca tctgcagcgc gctacccccc
      661 atcccgtacc acaagctcgc cgacctgcac tacctcagcc ggggcgcctc tggcacggtc
      721 tcgtcggcac gacacgcaga ctggcgcgtg cgggtggcag tgaagcacct acacatccac
      781 acgccgctgc tcgacagtga aagaaatgat atcttacgag aagccgaaat attacataaa
      841 gcaagattta gttacattct tccaattttg ggaatttgca atgagcctga atttttgggc
      901 atagttactg aatacatgcc aaatggatca ttaaatgagc ttctacacag aaaaactgaa
      961 tatcctgata ttgcatggcc attgagattc cgcatcctgc atgaaattgc tcttggtgta
     1021 aattatctgc acaatatgaa cccccctctg cttcatcatg acctaaagac tcagaatatc
     1081 ttgttggata atgaatttca tgttaagatt gcagattttg gtttatcaaa atggcgcatg
     1141 atgtcactct ctcaatcacg aagttacaaa tctgcacccg aaggaggaac aatcatctat
     1201 atgccacctg agaactatga gccaggacag aaatcaaggg ccagtgtgaa gcatgatata
     1261 tacagctatg cagttatcat gtgggaagtg ttatccagaa aacagccttt tgaagaggtc
     1321 accaatcctt tgcagatcat gtacagtgtg tcccagggac atcgacctga caccagtgag
     1381 gagaatttgc catttgatat acctcatcga ggtctcatga tctctctaat acagagtgga
     1441 tgggcgcaaa acccagatga aagaccatcc tttttgaaat gcttaataga acttgaacca
     1501 gttctgagaa catttgaaga cataactttc cttgaggctg ttattcagct aaagaaagca
     1561 aagatacaga gttcctcaag tactattcac ttatgtgata agaaaatgga tttatcgcta
     1621 aacatacctg caaatcatcc cccacaggag gaatcgtgtg gatcctctct gctctctaga
     1681 aacactggtt ctcctggacc ctccagatcc ctatcagctc ctcaagacaa gggattttta
     1741 tctggagcac ctcaagactg ctcttcgttg aaggcacacc actgtcctgg aaaccacagc
     1801 tgggatggta tcgtttctgt accaccaggg gcagccttct gtgatcgcag ggcctcctcg
     1861 tgttccttgg ctgtaataag tccattcttg gttgagaaag gttcagagcg cccaccaatt
     1921 ggtatagccc aacagtggat ccagagtaag agggaagcca ttgtgagcca gatgactgaa
     1981 gcctgcctta accaatcgct ggacgccctt ctgtctcgtg acctgatcat gaaggaggac
     2041 tatgaactca ttagcaccaa acctacaagg acctcaaaag tcagacagct gctagatacg
     2101 tctgacatcc aaggagaaga gtttgccaaa gttgtagtgc agaagttgaa agataacaag
     2161 cagctggggc tgcagcctta cccagaagta cctgtgcttt ccaaagcacc gccttcaaat
     2221 ttccctcaga ataaaagctt ggactacaaa gaccatgacg gtgattataa agatcatgac
     2281 atcgactaca aggatgacga tgacaagtag gaattctgca gtcgacggta ccgcgggccc
     2341 gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     2401 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     2461 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     2521 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2581 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2641 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2701 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     2761 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     2821 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     2881 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
     2941 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3001 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3061 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3121 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3181 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3241 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3301 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3361 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3421 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3481 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3541 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3601 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     3661 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3721 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3781 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3841 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3901 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3961 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4021 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4081 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4141 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4201 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4261 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4321 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4381 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4501 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4561 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4621 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4681 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4741 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4801 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4861 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4921 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4981 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5041 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     5101 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5161 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5221 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5281 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5341 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5401 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5461 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5521 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5581 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5641 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5701 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5761 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5821 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5881 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5941 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6001 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     6061 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6121 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6421 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6961 tctgtggata accgtattac cgccatgcat
//
