LOCUS       V031489                 8354 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031489
VERSION     V031489
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..3702
                     /label="CPED1(NM_024913)"
                     /note="CPED1(NM_024913)"
                     /gene="CPED1"
     misc_feature    3713..4299
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             4300..5019
                     /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    5142..5263
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5270..5725)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5752..5856
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        5858..6215
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6250..7044
                     /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    7276..7323
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      7652..8240
                     /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 caccatggtc tgtcgcccag tgttcccttg tcgtcggcga
      661 ttttgccccc gacccttctt ggtgggctta gtggtggcaa tctgtctctt ctaccagact
      721 ctgaccctcc gagggtcgag gaagctcaca gccgctgccc ctggggctgt cccacacaca
      781 tccactgaaa cccaggcaag cagatgcaag aaaggattct ctcaggacaa acagtgcttc
      841 cttctctctg gtaatgccca ggaaaccaga aaggtcaaag aatcaatgga gacacacttt
      901 ggcagccatg gccgaagggc catactctac aggcctcctt tctacagcaa aacagagctt
      961 cagctacacc agcacattct gactcaacat ggctatacgg ttgtcatcgc tgaagaaagg
     1021 ctcaatgctg gcctagggcc ggggctacta gaacaaggtg atctgggctc ttgggatctg
     1081 ctcatttgcc tgtcttctaa gaaagcagaa ggaacaccct gtatatccaa ggaagtcatg
     1141 tgccagttag gtttacatca aaaggcaaac agattaccag aaatacagca gccactttgc
     1201 agaaaggaag gattatgtca aatagttaga agattcccag aactgcaact tccagtgagt
     1261 ccctctgtgt gtctggatca gggaatgcaa ttaaagccga gtacttcgag tcacctttta
     1321 aaaacagtga agccacgtgt gtggaaacca ggggactgga gtcgtgaaca gctgaatgaa
     1381 acgacagtcc ttgctccaca tgaaacaatc tttcgagccg aagatctatc tgtgattctt
     1441 aaagcgtatg tgttggtgac gtccttaacc cctttgcgtg cattcattca ttcgacgggc
     1501 acagtttgga atccaccaaa gaaaaaacgc ttcactgtca agctgcaaac attttttgag
     1561 acattcctga gagccagttc acctcaacag gcttttgaca ttatgaagga agcaattggc
     1621 aaactactgc tagcggctga agtattcagt gaaacatcta ctctgggacc aaagaccttc
     1681 catagatgca gattctgctt tcaacttcta acttttgata ttggttatgg cagtttcatg
     1741 taccctgtag tgctccaggt acacgagcat ttaaattttc aagattatga taatatggat
     1801 tttgaggacc aaaatacaga agaattcctt ttaaatgaca ctttcaattt tctcttccct
     1861 aatgaatcat cactttccat attttctgag atatttcaga gactttatag atcagatgtt
     1921 ttcaagggtg aaaactatca aaaggaacta aatcagtgtc tgtccttaga agaaattaac
     1981 tcaattatga ctttcataaa ggaacttgga agtctgggac aattccaact gctcttccca
     2041 tctactactc ctgggattca gtcactgatg catgaatttt atgatgtggc aaatcctgtg
     2101 ggaaatcctg gctcagtcct gacccaatac tggtctcttt taaatgtatt tgaacaattt
     2161 cagttcatga ataaaaagac acagccacat ccactggaat ggaattcttt cacagaagat
     2221 aagaacattg aaaaaccaca agtgccattt gatgcaatag aaaataaaaa agctgcagtt
     2281 ccacaaatta aaaatgaaaa taaagaaata cattgcagtg atgatgaaaa cacaccatgt
     2341 catatcaagc agatcttcac acatccacat ttggaactaa atcctgactt tcatccaaag
     2401 atcaaagatt attactgtga agtcccattt gatgtggtaa cagtgacaat tggagtggaa
     2461 actcctaagt gtctgtgcaa ggtgcacctg tacgagcagg cagggccaag ctttgccagc
     2521 taccctctgg gcttaggaat gaacaaaatc tcaatatttg ttgtggatga atctccagca
     2581 cacggtgaga ctctgatcac gtacaaactc accatctata gagaagaccg cccaagtctg
     2641 cccttgtttg aggccttcac agcatgtggt tttgtgcagg attgtggttt gctgattcat
     2701 ccagaggaaa cctgtgggtt acagcctatt tcttctgact acattgaagc cattttacag
     2761 tctgaactaa aaagatgtcc atctggggac atgaaaggcc agtggattgt gccttgcctt
     2821 agttgttcgg acaacaggac gtgtgactgg agagaaataa cctggcagcc ccacaactgc
     2881 caatatggtg tcctaactaa gcctcaactc cagcagtgcc tgggaggaag gaagattctg
     2941 ttcattggag attcaaccaa cagagggatc atgtactatc ttattgaaag gctgaatgaa
     3001 acgttgcagg aatggcagaa agtacatggc actaaattct atcacaacgt caatggtggg
     3061 aagactttga tcagttattc ctactatccc cagttctgga taagcccttc attgagacca
     3121 acatttgaaa atgcacttga acacctcttg caaagatcac gtcccctaga gaatactggc
     3181 cagactgtat tggttgttgg tggtgttcag tggcttaatt ccaatcacct gcaaattatt
     3241 cacaaagttt tgaagaggga aaatttactc aatatcctag tgatcatcaa aactttggga
     3301 attggatttc atctgccagt ggatggagta catttcttaa cacagagtga agtacagaac
     3361 ttatggaaag aaaatttgat tattctggat actgcaaaaa aacatggcta tgaagtagtt
     3421 gacacattca ctataacaat ggggcgttac aaagagtttc tacaggggaa gtgtggatgt
     3481 catttccatg aggtagtgaa atcaaagtta tccaaagaat ataactttat taaaatgaaa
     3541 agatcaagaa atcatatcat gggaagatat ttcagcaatc aaagcaaact acaacaaggc
     3601 actgtaacaa attttcgatc gccatatcat gtcagaggtc caataaatca ggtttgttct
     3661 gaaatccttc tcagcaggat gtgtgcaaat aaaaggacta tgtagggatc cgcccctctc
     3721 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     3781 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     3841 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3901 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     3961 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4021 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     4081 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     4141 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     4201 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     4261 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     4321 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     4381 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     4441 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     4501 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4561 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4621 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4681 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4741 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4801 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4861 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4921 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4981 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     5041 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5101 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5161 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5221 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     5281 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5341 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5401 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5461 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5521 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5581 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5641 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5701 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5761 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5821 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5881 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5941 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     6001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     6061 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6121 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6181 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     6241 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6301 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6361 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6421 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6481 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6541 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6601 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6661 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6721 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     6781 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6841 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6901 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6961 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7021 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     7081 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7141 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7201 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     7261 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     7321 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     7381 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     7441 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     7501 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7561 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7621 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7681 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7741 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7801 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7861 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7981 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8041 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8101 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8161 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8221 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8281 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8341 ttaccgccat gcat
//
