LOCUS       V026791                 7700 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026791
VERSION     V026791
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..3190
                     /label="Creb5(NM_001327821)"
                     /note="Creb5(NM_001327821)"
                     /gene="Creb5"
     CDS             3197..3262
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3300..3524
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3570..3998
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4012..4341
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4408..5202
                     /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    5376..5497
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5546..5562)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5570..5586
                     /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(5594..5624)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5639..5660
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5948..6533)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6704..7564)
                     /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(7565..7669)
                     /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 aacttggagc aggagaggcc gtttgtctgc agtgccccag gctgctccca
     1741 gcgtttccca acagaggacc atctgatgat tcataggcac aagcacgaga tgactttgaa
     1801 gttcccttca ataaagacag acaatatgct gtcagatcag actccgacgc ccacaagatt
     1861 cctgaagaac tgtgaagagg tgggtctctt cagcgagctg gactgctcct ttgagcacga
     1921 gttcaggaag gctcaagagg aagagaacag caagcggaat atctccatgc ataatacagt
     1981 tggtgggacc atgacgggac ccggagctca ccagctaggc agcactcgga tgcccaacca
     2041 tgacagcagc gttgtgattc agcaagccat gccttctccc cagtccagct cggtcatcac
     2101 tcaggctcct tccaccaacc gccagattgg gcctgtccca ggctctctat catctctact
     2161 ccatctccac aacagacaga ggcagcccat gccggcctcc atgcctggaa ccctgcccaa
     2221 ccccaccatg ccaggatctt ctgccgtctt gatgcctatg gagagacaga tgtcagtgaa
     2281 ctccagcatc atgggcatgc aaggtccaaa cctcagcaac ccctgtgctt ctccccaagt
     2341 ccagccaatg cattcagaag ccaaaatgag actgaaggct gcgctgactc accatcctgc
     2401 cgccatgtcg aacgggaaca tgagcaccat cggacacatg atggagatga tgggctcccg
     2461 gcaagaccag acaccgcacc accacctgca ctcacacccg catcagcacc agacactgcc
     2521 gccccaccac ccctacccac accagcacca gcaccccgca caccatcccc acccacagcc
     2581 tcaccaccag cagaaccacc cgcaccacca ctcccattcc caccttcacg cacacccggc
     2641 gcaccaccag acctcgccac acccacccct gcccaccggc aaccaagcac aggtttcacc
     2701 agctacacaa cagatgcagc caacccagac aatacaaccg ccccagccca cagggggacg
     2761 ccggcgaaga gtggtggatg aggacccaga tgagaggcgg aggaaatttc tggaaaggaa
     2821 ccgggccgcc gccacccgct gcagacagaa gaggaaggtc tgggtgatgt cactggaaaa
     2881 gaaagctgaa gagctcaccc agacaaacat gcagcttcag aatgaagtgt ccatgttgaa
     2941 aaacgaggtg gcccagctga agcagttgtt gttaacacat aaagactgtc cgataacagc
     3001 catgcagaaa gaatcccaag ggtatttaag tccagagagc agccctcctg cgagccctgt
     3061 gccagcatgc tctcagcagc aagttatcca acacaacacc atcactacat cctcatcggt
     3121 cagcgaggtc gtggggagct ccaccctcag tcagcttaca actcacagaa cagacctgaa
     3181 tcctattctt ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     3241 ctacaaggat gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac
     3301 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3361 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3421 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3481 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3541 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3601 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3661 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3721 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3781 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3841 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3901 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3961 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4021 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4081 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4141 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4201 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4261 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4321 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4381 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4441 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4501 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4561 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4621 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4681 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4741 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4801 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4861 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4921 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4981 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5041 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5101 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5161 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5221 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5281 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5341 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5401 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5461 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5521 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5581 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5641 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5701 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6481 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctgacgtc
//
