LOCUS       V026523                 8618 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026523
VERSION     V026523
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             935..1651
                     /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             1688..4108
                     /label="OSBP(NM_002556)"
                     /note="OSBP(NM_002556)"
                     /gene="OSBP"
     CDS             4115..4180
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4218..4442
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4488..4916
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4930..5259
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5326..6120
                     /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    6294..6415
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6464..6480)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6488..6504
                     /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(6512..6542)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6557..6578
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6866..7451)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7622..8482)
                     /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(8483..8587)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccatggtg agcaagggcg aggagctgtt
      961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1141 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1621 gatcactctc ggcatggacg agctgtacaa ggaattcggt ggaggcggat caggaggcgg
     1681 tggatctatg gcggcgacgg agctgagagg agtggtgggg ccaggcccgg cagccattgc
     1741 agcacttggc ggcggcggcg ccggtccccc agtggtggga ggaggcggcg gccgcggaga
     1801 tgcggggcca ggctccgggg ccgcgtcagg gacggtggtc gcggcggcgg cgggaggccc
     1861 gggcccgggg gccgggggag tggcggcggc tggcccggcc cctgcgccgc cgactggggg
     1921 ctcgggcggc tcgggcgctg ggggttcggg ctcggctcga gagggctggc tcttcaaatg
     1981 gaccaattat atcaaaggct accagcggcg atggttcgtg ctgagcaacg ggctcctgag
     2041 ctactacaga tcaaaggcag aaatgagaca tacctgccgt ggtaccatca acctcgccac
     2101 agccaacatc accgtggagg actcctgcaa cttcatcatt tccaatgggg gtgctcagac
     2161 ctaccatctg aaagctagtt cagaagttga gcggcagcgc tgggtgacgg ccctggaact
     2221 ggccaaggcc aaagctgtga agatgctggc agagtcagat gaatcaggag atgaagagtc
     2281 tgtctcacaa actgacaaga ctgagctgca gaataccctt cggaccctct ctagcaaagt
     2341 agaggacttg agcacgtgca atgacttgat agctaagcat ggcacagctc tgcagcgttc
     2401 tctcagtgag ctggagtccc tgaagttgcc tgctgagagc aatgaaaaga tcaaacaggt
     2461 caacgaacga gccacactct ttaggataac atccaatgcc atgatcaacg cctgcagaga
     2521 tttcctcatg ttagcccaga cccatagtaa aaaatggcaa aagtcactac agtatgaaag
     2581 agaccagcgt atccgactgg aagaaaccct cgagcagctg gcgaagcagc ataatcacct
     2641 ggagagggcc ttccgaggag ccacggtgct gccggcaaac actcctggca atgtgggttc
     2701 tggtaaagat cagtgctgct ctggcaaagg ggacatgagc gatgaagatg atgagaatga
     2761 attttttgat gcacctgaga tcatcaccat gcctgaaaat ttgggccaca aacgtactgg
     2821 cagcaatatc agtggagcca gcagtgacat cagccttgat gaacagtaca agcatcagct
     2881 ggaggagacc aaaaaggaaa agagaaccag aataccatac aagccaaact atagcctcaa
     2941 tttatggagc atcatgaaga actgcattgg aaaagaactc tctaagatcc ccatgccggt
     3001 aaactttaat gagcccttgt ccatgcttca gcgccttact gaagatctgg aataccatga
     3061 gctgttagac cgagctgcaa aatgtgagaa ttctctagaa cagctctgtt atgttgcagc
     3121 tttcaccgtg tcctcctact ccactactgt cttccgcacc agtaagccat tcaacccact
     3181 gcttggggag acctttgagc tggaccgatt agaggagaat gggtaccgat ccctctgtga
     3241 acaggtgagt catcatcccc ctgctgctgc gcaccatgct gagtccaaaa atggctggac
     3301 attgcgtcag gaaatcaaaa tcaccagcaa gtttcgaggc aaatacctct ccattatgcc
     3361 cctcggtacc attcattgta ttttccatgc aactgggcac cactacactt ggaagaaagt
     3421 taccacaact gtacacaaca ttattgtggg caagttgtgg atagatcagt ctggcgaaat
     3481 tgatattgtg aatcacaaga caggagacaa gtgtaatctt aaatttgttc cttatagcta
     3541 cttctctcgg gatgtagcaa gaaaggtgac gggggaagtg acagatccat caggaaaagt
     3601 ccactttgct cttctgggga cgtgggatga gaaaatggaa tgtttcaaag tacagccagt
     3661 cattggggaa aatgggggtg atgctcgaca gagaggccat gaagcagagg aaagcagggt
     3721 catgctgtgg aaaaggaatc ctttaccgaa gaatgcagaa aacatgtact acttctcaga
     3781 gcttgctctg actctcaatg cttgggaaag tggcactgcc cccacagaca gccggttacg
     3841 acctgaccag agactgatgg aaaatggacg ctgggatgaa gcaaatgcgg agaagcagcg
     3901 cctggaggaa aaacaaagac tttccagaaa gaagagagaa gcggaagcta tgaaagccac
     3961 agaggatggc acaccatatg atccctataa ggcactgtgg tttgagcgga agaaggaccc
     4021 tgttaccaag gagttaaccc atatttatag gggagaatac tgggagtgta aagaaaaaca
     4081 ggactggagc tcatgcccgg acattttcct cgaggactac aaagaccatg acggtgatta
     4141 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     4201 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4261 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4321 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4381 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4441 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5161 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5281 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5341 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5401 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5461 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5521 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5581 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5641 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5701 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5761 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5821 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5881 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5941 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6001 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6061 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6121 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6181 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6241 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6301 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6361 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6421 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6481 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6541 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6601 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6661 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6721 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6781 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7381 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8581 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
