LOCUS       V022649                 6183 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022649
VERSION     V022649
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             613..1329
                     /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"
     CDS             1351..2838
                     /label="GSDME(NM_004403)"
                     /note="GSDME(NM_004403)"
                     /gene="GSDME"
     polyA_signal    2971..3092
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3099..3554)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3581..3685
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3687..4044
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4079..4873
                     /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    5105..5152
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5481..6069
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct atgtttgcca aagcaaccag gaattttctt
     1381 agagaagttg atgctgatgg tgacctgatt gcagtatcaa atctgaatga ctctgataag
     1441 ttacagcttc taagtctggt gacaaaaaag aagagattct ggtgctggca gagacccaag
     1501 taccagtttt tatccctcac ccttggcgat gtactcatag aagaccaatt tccgagtcca
     1561 gtggtcgtgg agtcggactt tgtgaaatac gagggcaagt ttgcaaacca cgtgagtgga
     1621 accctggaga ctgcactggg gaaggtcaag ctgaacctgg ggggcagcag ccgcgtagag
     1681 agccagtctt catttggaac cctgaggaag caggaggtgg atttgcagca gctcatcaga
     1741 gactctgccg agagaacaat aaatctgaga aaccctgtgc tccagcaggt gctggaagga
     1801 aggaatgagg tcctgtgcgt tttgacacag aagatcacga cgatgcagaa gtgtgtgatc
     1861 tctgagcaca tgcaggtcga ggagaagtgt ggtggcatcg tgggcatcca gaccaagacg
     1921 gtgcaggtgt cagcgacgga ggatgggaat gtcaccaagg actccaacgt ggtgctggag
     1981 atcccagctg ccaccaccat tgcctacggt gtcattgagt tatacgtgaa actggacggc
     2041 cagttcgagt tctgccttct ccgagggaag caaggtggct tcgagaacaa gaagagaatt
     2101 gactctgtct acctggaccc cctggtcttt cgagagtttg cattcataga catgccagat
     2161 gctgcgcatg ggatatcttc ccaggatgga ccattaagtg ttttaaagca agcgaccctg
     2221 ctcctggaga ggaatttcca tccatttgcg gagctgcctg agccacaaca gacagctttg
     2281 agtgacatct tccaggcggt cctatttgat gatgaactac tcatggtcct ggaaccagtg
     2341 tgcgatgacc tggtcagcgg cctctcgccc acagtggcgg tgctggggga gctgaagccc
     2401 cggcagcagc aggaccttgt ggccttcctg cagctggtgg ggtgcagctt acagggtggg
     2461 tgtccgggcc ccgaggatgc aggcagcaag cagctgttta tgacagccta cttcttggtc
     2521 agtgccctcg cagaaatgcc agatagcgca gcagctctgc tgggcacttg ctgcaaactc
     2581 cagatcattc ccacactgtg ccacttgctt cgtgctctgt ctgatgatgg agtatctgat
     2641 cttgaagacc caaccttgac tcccctgaaa gatacagaaa ggtttgggat tgtgcagcgc
     2701 ttgtttgcct cagctgacat tagtctggag agactgaagt catctgtgaa agctgtcatt
     2761 ctgaaggact ctaaagtctt cccactgctt ctttgtataa ccctgaatgg actctgtgct
     2821 ttaggcagag aacattcatg aggatccacc ggatctagat aactgatcat aatcagccat
     2881 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2941 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3061 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3121 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3181 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3241 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3301 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3361 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3421 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3481 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3541 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3661 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3721 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4081 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4141 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4201 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4261 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4321 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4381 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4441 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4501 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4561 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4621 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4681 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4741 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4801 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4861 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4921 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4981 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5041 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5101 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5161 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5221 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5281 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5341 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5401 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5461 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5521 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5581 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5641 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5701 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5761 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5821 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5881 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5941 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6001 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6061 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6121 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6181 cat
//
