LOCUS       V020001                 5764 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020001
VERSION     V020001
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             624..1676
                     /label="ATF4(NM_182810)"
                     /note="ATF4(NM_182810)"
                     /gene="ATF4"
     CDS             1710..2429
                     /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    2552..2673
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(2680..3135)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3162..3266
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3268..3625
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3660..4454
                     /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    4686..4733
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5062..5650
                     /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 atctcgagcc accatgaccg aaatgagctt cctgagcagc gaggtgttgg
      661 tgggggactt gatgtccccc ttcgaccagt cgggtttggg ggctgaagaa agcctaggtc
      721 tcttagatga ttacctggag gtggccaagc acttcaaacc tcatgggttc tccagcgaca
      781 aggctaaggc gggctcctcc gaatggctgg ctgtggatgg gttggtcagt ccctccaaca
      841 acagcaagga ggatgccttc tccgggacag attggatgtt ggagaaaatg gatttgaagg
      901 agttcgactt ggatgccctg ttgggtatag atgacctgga aaccatgcca gatgaccttc
      961 tgaccacgtt ggatgacact tgtgatctct ttgcccccct agtccaggag actaataagc
     1021 agccccccca gacggtgaac ccaattggcc atctcccaga aagtttaaca aaacccgacc
     1081 aggttgcccc cttcaccttc ttacaacctc ttcccctttc cccaggggtc ctgtcctcca
     1141 ctccagatca ttcctttagt ttagagctgg gcagtgaagt ggatatcact gaaggagata
     1201 ggaagccaga ctacactgct tacgttgcca tgatccctca gtgcataaag gaggaagaca
     1261 ccccttcaga taatgatagt ggcatctgta tgagcccaga gtcctatctg gggtctcctc
     1321 agcacagccc ctctaccagg ggctctccaa ataggagcct cccatctcca ggtgttctct
     1381 gtgggtctgc ccgtcccaaa ccttacgatc ctcctggaga gaagatggta gcagcaaaag
     1441 taaagggtga gaaactggat aagaagctga aaaaaatgga gcaaaacaag acagcagcca
     1501 ctaggtaccg ccagaagaag agggcggagc aggaggctct tactggtgag tgcaaagagc
     1561 tggaaaagaa gaacgaggct ctaaaagaga gggcggattc cctggccaag gagatccagt
     1621 acctgaaaga tttgatagaa gaggtccgca aggcaagggg gaagaaaagg gtccccggag
     1681 gagggggaag cgatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     1741 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1801 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1861 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1921 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1981 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2041 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2101 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2161 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2221 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2281 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     2341 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2401 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     2461 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2521 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2641 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     2701 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2761 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2821 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2881 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     2941 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3001 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3061 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3121 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3241 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3301 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3421 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3601 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3661 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3721 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3781 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3841 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3901 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3961 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4021 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4081 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4141 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4201 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4261 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4321 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4381 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4441 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4621 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4681 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4741 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4801 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4861 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     4921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     5761 gcat
//
