LOCUS       V041802                 9013 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041802
VERSION     V041802
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"
     regulatory      629..638
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             638..703
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             719..4141
                     /label="SREBF2(NM_004599)"
                     /note="SREBF2(NM_004599)"
                     /gene="SREBF2"
     polyA_signal    4182..4406
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     promoter        4422..4945
                     /label="mCMV promoter"
     CDS             4959..5678
                     /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    5801..5922
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5929..6384)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6411..6515
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        6517..6874
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6909..7703
                     /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    7935..7982
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      8311..8899
                     /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 atctcgagct ccgaattcgc caccatggac tacaaagacc atgacggtga
      661 ttataaagat catgacatcg actacaagga tgacgatgac aagggtggag gcggatcaat
      721 ggacgacagc ggcgagctgg gtggtctgga gaccatggag accctcacgg agctgggcga
      781 cgagctgacc ctgggagaca tcgacgagat gctgcaattt gtcagtaatc aagtgggaga
      841 gttccctgac ttgttttcag aacagctgtg tagctccttt cctggcagtg gtggtagtgg
      901 tagcagcagc ggcagcagtg gcagcagcag cagcagcagc aatggcaggg gcagcagcag
      961 cggagctgtg gacccttcag tgcaacggtc attcacccag gtcacattac cttccttctc
     1021 tccctcggcg gcctccccac aggctccaac tctgcaagtc aaggtttctc ccacctcagt
     1081 tcccaccaca cccagggcaa ctcctattct tcagccccgc ccccagcccc agcctcaacc
     1141 tcaaactcag ctgcaacaac agacggtaat gatcacgcca acattcagca ccactccgca
     1201 gacgaggatc atccagcagc ctttgatata ccagaatgca gctactagct ttcaagtcct
     1261 tcagcctcaa gtccaaagcc tggtgacatc ctcccaggta cagccggtca ccattcagca
     1321 gcaggtgcag acagtacagg cccagcgggt gctgacacaa acggccaatg gcacgctgca
     1381 gacccttgcc ccggctacgg tgcagacagt tgctgcgcca caggtgcagc aggtcccggt
     1441 cctggtccag cctcagatca tcaagacaga ttcccttgtt ttgaccacac tgaagacaga
     1501 tggcagccct gttatggctg cggtccagaa cccggccctc accgccctca ccacccctat
     1561 ccagacggct gcccttcaag taccaaccct ggtgggcagc agtgggacca ttctgaccac
     1621 aatgcctgta atgatggggc aagagaaagt gcccattaag caggtacctg ggggagtcaa
     1681 gcagcttgag ccccccaaag aaggagaaag gcggacaacc cataatatca ttgagaaacg
     1741 atatcgctcc tccatcaatg acaaaatcat cgaattgaaa gacctggtca tggggacaga
     1801 cgccaagatg cacaagtctg gcgttctgag gaaggccatt gattacatca aatacttgca
     1861 gcaggtcaat cataaactgc gccaggagaa catggtgctg aagctggcaa atcaaaagaa
     1921 caagcttcta aagggcatcg acctaggcag tctggtggac aatgaggtgg acctgaagat
     1981 cgaggacttt aatcagaatg tccttctgat gtccccccca gcctctgact cagggtccca
     2041 ggctggcttc tctccctact ccattgactc tgagccagga agccctctat tggatgatgc
     2101 aaaggtcaaa gatgagccag actctcctcc tgtggcgctg ggcatggtag accgctcacg
     2161 gattcttctg tgtgtcctca ccttcctgtg cctctccttt aaccccctga cttccctgct
     2221 gcagtgggga ggggcccacg actctgacca gcacccacac tcaggctctg gccgcagtgt
     2281 cctgtcattc gagtcaggtt ctgggggctg gtttgactgg atgatgccta ctcttctctt
     2341 atggctggta aatggtgtga ttgtcctgag cgtctttgtg aagctgctgg ttcatgggga
     2401 gccagtgatc cggccacact cgcgctcctc ggtcaccttc tggaggcacc ggaaacaggc
     2461 agatctggat ctcgccagag gagattttgc agctgctgcc ggcaacctac aaacctgcct
     2521 ggcagttttg ggccgggcac tgcccacctc ccgcctggac ctggcctgca gcctctcctg
     2581 gaacgtgatc cgctacagcc tgcagaagct acgcctggtg cgctggctgc tcaagaaagt
     2641 cttccagtgc cggcgggcca cgccagccac tgaggcaggc tttgaagacg aagctaagac
     2701 cagcgcccgg gatgcggctc tggcctatca ccggctgcac cagctgcaca tcacagggaa
     2761 gcttcctgca ggatccgcct gttccgatgt acacatggcg ttgtgtgccg tgaacctggc
     2821 tgaatgtgca gaggagaaga tcccaccgag cacactggtt gagatccatc tgactgctgc
     2881 catggggctc aagacccggt gtggaggcaa gctgggcttc ctggccagct acttcctcag
     2941 ccgagcccag agcctgtgtg gccccgagca cagtgctgtt cctgactccc tgcgctggct
     3001 ctgccacccc ctgggccaga agtttttcat ggagcggagc tggtctgtga agtcagctgc
     3061 caaggagagt ctatactgtg cccagaggaa cccagctgac cccattgcgc aggtccacca
     3121 ggccttctgc aagaacctgc tggagcgagc tatagagtcc ttggtgaaac ctcaggccaa
     3181 gaagaaggct ggagaccagg aagaagagag ctgtgaattc tccagtgctc tggagtactt
     3241 gaaattactt cattcttttg tggactctgt gggggttatg agccccccac tctccaggag
     3301 ctccgtgctc aagtccgccc tgggtccaga catcatctgt cggtggtgga cgtctgcaat
     3361 cactgtggcc atcagctggc tccagggaga cgatgcagct gtgcgctctc attttaccaa
     3421 agtggaacgc atccccaagg ccctggaagt gacagagagc cccctggtga aggccatctt
     3481 ccatgcctgc agagccatgc atgcctcact ccctgggaaa gcagatgggc agcagagttc
     3541 cttctgccat tgcgagaggg ccagtggcca cctatggagc agcctcaacg tcagtggggc
     3601 cacctctgac cctgccctca accacgtggt ccagctgctc acctgtgacc tgctactgtc
     3661 gctacggaca gcgctctggc aaaaacaggc cagtgccagc caggctgtgg gggagaccta
     3721 ccacgcgtca ggcgctgaac tggcgggctt ccaacgggac ctgggcagcc tgcgcaggct
     3781 ggcacacagc ttccgcccag cataccgcaa ggtgttcctg catgaagcca ccgtgcgcct
     3841 gatggcagga gccagcccca cccgcaccca ccagctgctg gaacacagcc tgcggcggcg
     3901 caccacgcag agcaccaagc acggagaggt ggatgcctgg cccggccagc gagagcgggc
     3961 caccgccatc ctgctggcct gccgccacct gcccctctcc ttcctctcct ccccgggcca
     4021 gcgggcagtg ctgctggccg aagctgcccg caccctggag aaggtgggcg accggcgctc
     4081 ctgcaacgac tgccagcaga tgattgttaa gctgggtggt ggcactgcca ttgccgcctc
     4141 ctgaggatcc gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4201 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4261 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4321 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4381 tgctggggat gcggtgggct ctatggtcta gagatatact gagtcattag ggactttcca
     4441 atgggttttg cccagtacat aaggtcaata ggggtgaatc aacaggaaag tcccattgga
     4501 gccaagtaca ctgagtcaat agggactttc cattgggttt tgcccagtac aaaaggtcaa
     4561 tagggggtga gtcaatgggt ttttcccatt attggcacgt acataaggtc aataggggtg
     4621 agtcattggg tttttccagc caatttaatt aaaacgccat gtactttccc accattgacg
     4681 tcaatgggct attgaaacta atgcaacgtg acctttaaac ggtactttcc catagctgat
     4741 taatgggaaa gtaccgttct cgagccaata cacgtcaatg ggaagtgaaa gggcagccaa
     4801 aacgtaacac cgccccggtt ttcccctgga aattccatat tggcacgcat tctattggct
     4861 gagctgcgtt ctacgtgggt ataagaggcg cgaccagcgt cggtaccgtc gcagtcttcg
     4921 gtctgaccac cgtagaacgc agatcaccgg tcgccaccat ggtgagcaag ggcgaggagc
     4981 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     5041 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     5101 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     5161 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     5221 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     5281 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     5341 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     5401 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     5461 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     5521 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     5581 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     5641 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     5701 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5761 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5821 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5881 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     5941 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6001 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6061 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6121 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6181 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6241 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6301 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6361 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6421 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6481 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     6541 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6601 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     6661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     6721 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6781 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6841 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     6901 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     6961 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     7021 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     7081 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     7141 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     7201 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     7261 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     7321 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     7381 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     7441 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     7501 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     7561 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     7621 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     7681 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     7741 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7801 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7861 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     7921 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     7981 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     8041 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     8101 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     8161 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8221 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8281 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     8341 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8401 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     8461 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8521 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8581 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8641 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     8701 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8761 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8821 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8881 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8941 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     9001 taccgccatg cat
//
