LOCUS       V034027                 7637 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034027
VERSION     V034027
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             923..1639
                     /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             1676..3127
                     /label="Osbpl2(NM_144500)"
                     /note="Osbpl2(NM_144500)"
                     /gene="Osbpl2"
     CDS             3134..3199
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3237..3461
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3507..3935
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3949..4278
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4345..5139
                     /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    5313..5434
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5483..5499)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5507..5523
                     /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(5531..5561)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5576..5597
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5885..6470)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6641..7501)
                     /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(7502..7606)
                     /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 aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatgaa
     1681 cggggaggaa gaattctttg atgccgtcac aggctttgat tctgataatt cttctatagg
     1741 ggaattttca gaggcaaata aaattagtgg gatgattgac ttggatacca gcaaaagtac
     1801 taggagtggg aaaaatgggg agaaaccaca acaagagaat ggaattcaaa agcacaggac
     1861 agcccttcct gctcccatgt tcaccagaag cgacttcagt gtgtggagta tcctgaagaa
     1921 atgcattggc ttggaactct ccaagatcac aatgccaatt gccttcaatg agccactgag
     1981 cttcctgcag cggatcacag agtacatgga gcatgtgtac ctcattcaca aagcctcaag
     2041 tcagtcccag ccgctggaga gaatgcagtc tgtcgctgcc tttgcggtct cagctgtggc
     2101 ttcccagtgg gagagaactg gcaaaccctt taacccactc ttgggagaga catatgagtt
     2161 gatcagggaa gacttagggt tcagatttat atctgagcag gtcagtcacc accctcctat
     2221 tagcgcattt tattcagaag gtctcaatca agacttcagg ttccacggtt ccatctaccc
     2281 aaagctgaag ttctggggca agagtgtgga ggcggagccc cgggggacca tcacactgga
     2341 gctgctcaaa cataatgaag cctatacctg gaccaaccct acctgctgtg tacacaatgt
     2401 catccttggg cagctatgga tagagcagta tgggatagtg gagattgtaa atcacagaac
     2461 tggagataag tgtatacttc acttcaagcc ttgtggattg tttgggaagg aacttcacag
     2521 agtagaaggg tacattcaag acaaaaacag gaagaagctc tttatcatgt atggcaagtg
     2581 gacagagtgc ttgtggggaa tagatcctgc ttcatacgag tcctttaaga agcaagagaa
     2641 gagaggtgac caggccagga aggccaagat ggatgatggc cccgagaaag ctaacagtga
     2701 tgtgcctggt gatgtggcag acgacgttcc tgtggctcag gagacggtgc aagtcatccc
     2761 cggcagcaag ctgctctgga ggatcaacag ccggcccccc aactctgctc agatgtacaa
     2821 ctttaccagc ttcactgtga gtctcaatga gttggagtca ggcatggaga agactctgcc
     2881 acccacagac tgccgcctcc gtcctgacat acgaggcatg gagaacggaa acatggatct
     2941 ggccagccag gagaaggagc ggctagagga aaagcagaga gaggccagaa aagagcgtgc
     3001 caaagaggac gcagagtggc ggaccaggtg gttctctcca ggcaataacc cctacacagg
     3061 agctcctgac tggctgtacg ccggccatta ctttgagcgt aacttctcag actgcccaga
     3121 catatacctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3181 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     3241 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3301 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3361 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3421 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3481 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3541 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3601 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3661 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3721 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3781 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3841 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3901 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4321 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4381 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4441 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4501 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4561 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4621 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4681 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4741 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4801 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4861 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4921 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4981 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5041 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5101 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5161 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5221 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5281 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5341 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5401 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5461 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5521 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5581 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5641 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5701 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6421 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6481 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6541 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6601 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6661 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6721 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6781 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6841 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6901 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6961 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7021 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7081 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7141 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7201 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7261 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7321 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7381 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7441 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7501 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7561 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7621 aagtgccacc tgacgtc
//
