LOCUS       V043116                 8083 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043116
VERSION     V043116
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             937..1653
                     /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             1690..3834
                     /label="FOXP2(NM_014491)"
                     /note="FOXP2(NM_014491)"
                     /gene="FOXP2"
     CDS             3847..3912
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3947..4171
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4217..4645
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4659..4988
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5000..5599
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5759..5880
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5929..5945)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5953..5969
                     /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(5977..6007)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6022..6043
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6331..6916)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7087..7947)
                     /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(7948..8052)
                     /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 aagctttcta gagaattcga tatcggatcc gccaccatgg tgagcaaggg cgaggagctg
      961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1621 gggatcactc tcggcatgga cgagctgtac aagggcggtg gaggcagtgg tggaggcggt
     1681 agcggtacca tgatgcagga atctgcgaca gagacaataa gcaacagttc aatgaatcaa
     1741 aatggaatga gcactctaag cagccaatta gatgctggca gcagagatgg aagatcaagt
     1801 ggtgacacca gctctgaagt aagcacagta gaactgctgc atctgcaaca acagcaggct
     1861 ctccaggcag caagacaact tcttttacag cagcaaacaa gtggattgaa atctcctaag
     1921 agcagtgata aacagagacc actgcaggtg cctgtgtcag tggccatgat gactccccag
     1981 gtgatcaccc ctcagcaaat gcagcagatc cttcagcaac aagtcctgtc tcctcagcag
     2041 ctacaagccc ttctccaaca acagcaggct gtcatgctgc agcagcaaca actacaagag
     2101 ttttacaaga aacagcaaga gcagttacat cttcagcttt tgcagcagca gcagcaacag
     2161 cagcagcagc aacaacagca gcaacaacag cagcagcaac aacaacaaca acagcagcaa
     2221 caacagcagc agcagcagca acagcagcag cagcagcaac agcatcctgg aaagcaagcg
     2281 aaagagcagc agcagcagca gcagcagcaa cagcaattgg cagcccagca gcttgtcttc
     2341 cagcagcagc ttctccagat gcaacaactc cagcagcagc agcatctgct cagccttcag
     2401 cgtcagggac tcatctccat tccacctggc caggcagcac ttcctgtcca atcgctgcct
     2461 caagctggct taagtcctgc tgagattcag cagttatgga aagaagtgac tggagttcac
     2521 agtatggaag acaatggcat taaacatgga gggctagacc tcactactaa caattcctcc
     2581 tcgactacct cctccaacac ttccaaagca tcaccaccaa taactcatca ttccatagtg
     2641 aatggacagt cttcagttct aagtgcaaga cgagacagct cgtcacatga ggagactggg
     2701 gcctctcaca ctctctatgg ccatggagtt tgcaaatggc caggctgtga aagcatttgt
     2761 gaagattttg gacagttttt aaagcacctt aacaatgaac acgcattgga tgaccgaagc
     2821 actgctcagt gtcgagtgca aatgcaggtg gtgcaacagt tagaaataca gctttctaaa
     2881 gaacgcgaac gtcttcaagc aatgatgacc cacttgcaca tgcgaccctc agagcccaaa
     2941 ccatctccca aacctctaaa tctggtgtct agtgtcacca tgtcgaagaa tatgttggag
     3001 acatccccac agagcttacc tcaaacccct accacaccaa cggccccagt caccccgatt
     3061 acccagggac cctcagtaat caccccagcc agtgtgccca atgtgggagc catacgaagg
     3121 cgacattcag acaaatacaa cattcccatg tcatcagaaa ttgccccaaa ctatgaattt
     3181 tataaaaatg cagatgtcag acctccattt acttatgcaa ctctcataag gcaggctatc
     3241 atggagtcat ctgacaggca gttaacactt aatgaaattt acagctggtt tacacggaca
     3301 tttgcttact tcaggcgtaa tgcagcaact tggaagaatg cagtacgtca taatcttagc
     3361 ctgcacaagt gttttgttcg agtagaaaat gttaaaggag cagtatggac tgtggatgaa
     3421 gtagaatacc agaagcgaag gtcacaaaag ataacaggaa gtccaacctt agtaaaaaat
     3481 atacctacca gtttaggcta tggagcagct cttaatgcca gtttgcaggc tgccttggca
     3541 gagagcagtt tacctttgct aagtaatcct ggactgataa ataatgcatc cagtggccta
     3601 ctgcaggccg tccacgaaga cctcaatggt tctctggatc acattgacag caatggaaac
     3661 agtagtccgg gctgctcacc tcagccgcac atacattcaa tccacgtcaa ggaagagcca
     3721 gtgattgcag aggatgaaga ctgcccaatg tccttagtga caacagctaa tcacagtcca
     3781 gaattagaag acgacagaga gattgaagaa gagcctttat ctgaagatct ggaactcgag
     3841 accggtgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     3901 gacgatgaca agtgaccggt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     3961 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4021 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4081 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4141 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4201 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4561 tattcttttg atttataagg gattttggcc atttcggcct attggttaaa aaatgagctg
     4621 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4681 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4921 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4981 tttgcaaaaa gctcccggga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5041 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5101 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5161 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5221 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5281 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5341 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5401 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5461 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5521 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5581 ccgcaagccc ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5641 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5701 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5761 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5821 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5881 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5941 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6001 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6061 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6841 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7081 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7141 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7201 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7261 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7321 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7381 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7441 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7501 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7561 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7621 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7681 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7741 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7801 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7861 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7921 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7981 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8041 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
