LOCUS       V032267                 6511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032267
VERSION     V032267
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             635..1561
                     /label="Ucp2(NM_011671)"
                     /note="Ucp2(NM_011671)"
                     /gene="Ucp2"
     CDS             1574..1639
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1680..1904
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     promoter        1920..2443
                     /label="mCMV promoter"
     CDS             2457..3176
                     /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    3299..3420
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3427..3882)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3909..4013
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4015..4372
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4407..5201
                     /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    5433..5480
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5809..6397
                     /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 caccatggtt ggtttcaagg ccacagatgt
      661 gcccccaaca gccactgtga agttcctggg ggctgggaca gctgcctgca ttgcagatct
      721 catcactttc cctctggata ccgccaaggt ccggctgcag atccaagggg agagtcaagg
      781 gctagtgcgc accgcagcca gcgcccagta ccgtggcgtt ctgggtacca tcctaaccat
      841 ggtgcgcact gagggtccac gcagcctcta caatgggctg gtcgccggcc tgcagcgcca
      901 gatgagcttt gcctccgtcc gcattggcct ctacgactct gtcaaacagt tctacaccaa
      961 gggctcagag catgcaggca tcgggagccg cctcctggca ggtagcacca caggtgccct
     1021 ggccgtggct gtagcccagc ctacagatgt ggtaaaggtc cgcttccagg ctcaggcccg
     1081 ggctggtggt ggtcggagat accagagcac tgtcgaagcc tacaagacca ttgcacgaga
     1141 ggaagggatc cggggcctct ggaaagggac ttctcccaat gttgcccgta atgccattgt
     1201 caactgtgct gagctggtga cctatgacct catcaaagat actctcctga aagccaacct
     1261 catgacagat gacctccctt gccacttcac ttctgccttc ggggccggct tctgcaccac
     1321 cgtcatcgcc tcccctgttg atgtggtcaa gacgagatac atgaactctg ccttgggcca
     1381 gtaccacagc gcaggtcact gtgcccttac catgctccgg aaggagggac cccgcgcctt
     1441 ctacaagggg ttcatgcctt cctttctccg cttgggatcc tggaacgtag tgatgtttgt
     1501 cacctatgag cagctcaaaa gagccctaat ggctgcctac caatctcggg aggcaccttt
     1561 caagcttacg cgtgactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     1621 caaggatgac gatgacaagt gaggatccgg gcccgtttaa acccgctgat cagcctcgac
     1681 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1741 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1801 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1861 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggtctaga gatatactga
     1921 gtcattaggg actttccaat gggttttgcc cagtacataa ggtcaatagg ggtgaatcaa
     1981 caggaaagtc ccattggagc caagtacact gagtcaatag ggactttcca ttgggttttg
     2041 cccagtacaa aaggtcaata gggggtgagt caatgggttt ttcccattat tggcacgtac
     2101 ataaggtcaa taggggtgag tcattgggtt tttccagcca atttaattaa aacgccatgt
     2161 actttcccac cattgacgtc aatgggctat tgaaactaat gcaacgtgac ctttaaacgg
     2221 tactttccca tagctgatta atgggaaagt accgttctcg agccaataca cgtcaatggg
     2281 aagtgaaagg gcagccaaaa cgtaacaccg ccccggtttt cccctggaaa ttccatattg
     2341 gcacgcattc tattggctga gctgcgttct acgtgggtat aagaggcgcg accagcgtcg
     2401 gtaccgtcgc agtcttcggt ctgaccaccg tagaacgcag atcaccggtc gccaccatgg
     2461 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2521 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2581 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2641 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2701 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2761 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2821 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2881 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2941 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3001 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3061 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3121 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     3181 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3241 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3301 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3361 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3421 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3481 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3541 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3601 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3661 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3721 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3781 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3841 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4021 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4381 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4441 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4501 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4561 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4621 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4681 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4741 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4801 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4861 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4921 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4981 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5041 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5101 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5161 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5221 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5281 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5341 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5401 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5461 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5521 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5581 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5641 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5941 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6421 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6481 ttctgtggat aaccgtatta ccgccatgca t
//
