LOCUS       V031576                 8594 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031576
VERSION     V031576
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             1658..4084
                     /label="PDE4D(NM_001104631)"
                     /note="PDE4D(NM_001104631)"
                     /gene="PDE4D"
     CDS             4091..4156
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4194..4418
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4464..4892
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4906..5235
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5302..6096
                     /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    6270..6391
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6440..6456)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6464..6480
                     /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(6488..6518)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6533..6554
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6842..7427)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7598..8458)
                     /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(8459..8563)
                     /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 ggaattcatg gaggcagagg gcagcagcgc
     1681 gccggcccgg gcgggcagcg gagagggcag cgacagcgcc ggcggggcca cgctcaaagc
     1741 ccccaagcat ctctggaggc acgagcagca ccaccagtac ccgctccggc agccccagtt
     1801 ccgcctcctg catccccatc accacctgcc cccgccgccg ccaccctcgc cccagcccca
     1861 gccccagtgt ccgctacagc cgccgccgcc gccccccctg ccgccgcccc cgccgccgcc
     1921 cggggctgcc cgcggccgct acgcctcgag cggggccacc ggccgcgtcc ggcatcgcgg
     1981 ctactcggac accgagcgct acctgtactg tcgcgccatg gaccgcacct cctacgcggt
     2041 ggagaccggc caccggcccg gcctgaagaa atccaggatg tcctggccct cctcgttcca
     2101 gggactcagg cgttttgatg tggacaatgg cacatctgcg ggacggagtc ccttggatcc
     2161 catgaccagc ccaggatccg ggctaattct ccaagcaaat tttgtccaca gtcaacgacg
     2221 ggagtccttc ctgtatcgat ccgacagcga ttatgacctc tctccaaagt ctatgtcccg
     2281 gaactcctcc attgccagtg atatacacgg agatgacttg attgtgactc catttgctca
     2341 ggtcttggcc agtctgcgaa ctgtacgaaa caactttgct gcattaacta atttgcaaga
     2401 tcgagcacct agcaaaagat cacccatgtg caaccaacca tccatcaaca aagccaccat
     2461 aacagaggag gcctaccaga aactggccag cgagaccctg gaggagctgg actggtgtct
     2521 ggaccagcta gagaccctac agaccaggca ctccgtcagt gagatggcct ccaacaagtt
     2581 taaaaggatg cttaatcggg agctcaccca tctctctgaa atgagtcggt ctggaaatca
     2641 agtgtcagag tttatatcaa acacattctt agataagcaa catgaagtgg aaattccttc
     2701 tccaactcag aaggaaaagg agaaaaagaa aagaccaatg tctcagatca gtggagtcaa
     2761 gaaattgatg cacagctcta gtctgactaa ttcaagtatc ccaaggtttg gagttaaaac
     2821 tgaacaagaa gatgtccttg ccaaggaact agaagatgtg aacaaatggg gtcttcatgt
     2881 tttcagaata gcagagttgt ctggtaaccg gcccttgact gttatcatgc acaccatttt
     2941 tcaggaacgg gatttattaa aaacatttaa aattccagta gatactttaa ttacatatct
     3001 tatgactctc gaagaccatt accatgctga tgtggcctat cacaacaata tccatgctgc
     3061 agatgttgtc cagtctactc atgtgctatt atctacacct gctttggagg ctgtgtttac
     3121 agatttggag attcttgcag caatttttgc cagtgcaata catgatgtag atcatcctgg
     3181 tgtgtccaat caatttctga tcaatacaaa ctctgaactt gccttgatgt acaatgattc
     3241 ctcagtctta gagaaccatc atttggctgt gggctttaaa ttgcttcagg aagaaaactg
     3301 tgacattttc cagaatttga ccaaaaaaca aagacaatct ttaaggaaaa tggtcattga
     3361 catcgtactt gcaacagata tgtcaaaaca catgaatcta ctggctgatt tgaagactat
     3421 ggttgaaact aagaaagtga caagctctgg agttcttctt cttgataatt attccgatag
     3481 gattcaggtt cttcagaata tggtgcactg tgcagatctg agcaacccaa caaagcctct
     3541 ccagctgtac cgccagtgga cggaccggat aatggaggag ttcttccgcc aaggagaccg
     3601 agagagggaa cgtggcatgg agataagccc catgtgtgac aagcacaatg cttccgtgga
     3661 aaaatcacag gtgggcttca tagactatat tgttcatccc ctctgggaga catgggcaga
     3721 cctcgtccac cctgacgccc aggatatttt ggacactttg gaggacaatc gtgaatggta
     3781 ccagagcaca atccctcaga gcccctctcc tgcacctgat gacccagagg agggccggca
     3841 gggtcaaact gagaaattcc agtttgaact aactttagag gaagatggtg agtcagacac
     3901 ggaaaaggac agtggcagtc aagtggaaga agacactagc tgcagtgact ccaagactct
     3961 ttgtactcaa gactcagagt ctactgaaat tccccttgat gaacaggttg aagaggaggc
     4021 agtaggggaa gaagaggaaa gccagcctga agcctgtgtc atagatgatc gttctcctga
     4081 cacgctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     4141 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     4201 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4261 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4321 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4381 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4441 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4501 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4561 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4621 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4681 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4741 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4801 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4861 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4921 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4981 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5041 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5101 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5161 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5221 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5281 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5341 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5401 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5461 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5521 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5581 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5641 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5701 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5761 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5821 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5881 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5941 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6001 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6061 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6121 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6181 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6241 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6301 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6361 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6421 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6481 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6541 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7261 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     7381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7501 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7561 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7681 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7741 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7801 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8041 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8101 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8281 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8521 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8581 tgccacctga cgtc
//
