LOCUS       V041377                 7783 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041377
VERSION     V041377
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             966..1682
                     /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             1734..3284
                     /label="TRIM65(NM_173547)"
                     /note="TRIM65(NM_173547)"
                     /gene="TRIM65"
     CDS             3285..3350
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3383..3607
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3653..4081
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4095..4424
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4491..5285
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5459..5580
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5629..5645)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5653..5669
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5677..5707)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5722..5743
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6031..6616)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6787..7647)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7648..7752)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcg
      961 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1021 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1081 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1141 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1201 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1261 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1321 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1381 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1441 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1501 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1561 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1621 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1681 agggaggcgg aggtagtgga ggagggggaa gcggaggcgg agggagtgaa ttcatggccg
     1741 cgcagctgct ggaggagaag ctgacctgcg ccatctgcct ggggctctac caggacccag
     1801 tgacgctgcc ctgcggccac aacttctgcg gggcctgcat ccgggactgg tgggaccgct
     1861 gcggaaaggc gtgccccgag tgccgggagc cctttcccga cggcgccgag ctgcgccgca
     1921 acgtggccct cagcggcgtg ctggaggtgg tgcgcgccgg gcccgcccgg gatcccggcc
     1981 ccgatcccgg ccccggcccc gaccctgccg cgcgctgccc ccgccacggg cggccgctgg
     2041 agctcttctg ccggaccgag ggccgctgtg tgtgcagcgt gtgcaccgtg cgcgagtgtc
     2101 gcctccacga gcgggcgctg ctggatgccg agcgcctcaa gcgcgaggcc cagctgagag
     2161 ccagcctgga ggttacccag cagcaggcca cccaggccga aggccagcta ctagagctgc
     2221 gcaagcaaag cagccagatc cagaactcgg cctgcatctt ggcctcctgg gtctccggca
     2281 agttcagcag cctgctacag gccctggaaa tacagcacac gacagcactg aggagcatcg
     2341 aggtggccaa gacgcaggcg ctggcacagg ctcgagacga ggagcagcgg ctgcgggtcc
     2401 atttggaggc tgtggctcgc catggctgca ggatccggga gctcctggag caggtggatg
     2461 agcagacctt cctgcaggaa tcgcagctcc tccagccccc agggcctctt gggccactga
     2521 cccctctgca gtgggatgaa gaccaacagc tgggtgacct gaagcagttg ctaagccggc
     2581 tgtgtggcct cctcttggaa gaggggagcc accctggggc accagccaag cctgtggact
     2641 tagcccccgt ggaggcccca ggtcccctgg caccggtccc aagcacagtt tgtccactga
     2701 ggaggaaact ctggcagaat tatcgcaatc tgacctttga tccagtcagc gccaaccgtc
     2761 acttctatct gtcgcgccag gaccagcagg tgaagcactg tcgtcagtcc cggggcccag
     2821 gcgggcccgg cagctttgag ctctggcagg tgcaatgtgc ccagagcttc caggccgggc
     2881 accactactg ggaggtgcgc gcgtcagacc actcggtgac actgggcgtc tcctacccgc
     2941 aactgccacg gtgcaggctg gggccccaca cagacaacat tggccgggga ccctgctcct
     3001 gggggctctg cgtccaggag gacagcctcc aggcctggca caacggggaa gcccagcgcc
     3061 tcccaggggt gtcagggcgg ctcctgggca tggatttgga cctggcctca ggctgcctca
     3121 ccttctacag cctggagccc cagacccagc ccctgtacac cttccatgcc ctcttcaacc
     3181 agcccctcac ccccgtcttc tggctcctcg agggtaggac cctgaccctg tgccatcagc
     3241 caggggctgt gttccctctg gggccccagg aagaggtgct cagcgactac aaagaccatg
     3301 acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag tagcttaagt
     3361 ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3421 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3481 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3541 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3601 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3841 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3961 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4021 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4081 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4141 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4201 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4261 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4321 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4381 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4441 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4501 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4561 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4621 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4681 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4741 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4801 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4861 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4921 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4981 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5041 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5101 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5161 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5221 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5281 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5341 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5401 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5461 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5521 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5581 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5641 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5701 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5761 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5821 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6541 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6601 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6661 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6721 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6781 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6841 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6901 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6961 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7021 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7081 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7141 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7201 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7261 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7321 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7381 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7441 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7501 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7561 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7621 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7681 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7741 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
