LOCUS       V039627                 8333 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039627
VERSION     V039627
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..3823
                     /label="IKBKE(NM_014002)"
                     /note="IKBKE(NM_014002)"
                     /gene="IKBKE"
     CDS             3830..3895
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3933..4157
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4203..4631
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4645..4974
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5041..5835
                     /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    6009..6130
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6179..6195)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6203..6219
                     /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(6227..6257)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6272..6293
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6581..7166)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7337..8197)
                     /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(8198..8302)
                     /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 aattcatgca
     1681 gagcacagcc aattacctgt ggcacacaga tgacctgctg gggcaggggg ccactgccag
     1741 tgtgtacaag gcccgcaaca agaaatccgg agagctggtt gctgtgaagg tcttcaacac
     1801 taccagctac ctgcggcccc gcgaggtgca ggtgagggag tttgaggtcc tgcggaagct
     1861 gaaccaccag aacattgtca agctctttgc ggtggaggag acgggcggaa gccggcagaa
     1921 ggtactggtg atggagtact gctccagtgg gagcctgctg agtgtgctgg agagccctga
     1981 gaatgccttt gggctgcctg aggatgagtt cctggtggtg ctgcgctgtg tggtggccgg
     2041 catgaaccac ctgcgggaga acggcattgt gcatcgcgac atcaagccgg ggaacatcat
     2101 gcgcctcgta ggggaggagg ggcagagcat ctacaagctg acagacttcg gcgctgcccg
     2161 ggagctggat gatgatgaga agttcgtctc ggtctatggg actgaggagt acctgcatcc
     2221 cgacatgtat gagcgggcgg tgcttcgaaa gccccagcaa aaagcgttcg gggtgactgt
     2281 ggatctctgg agcattggag tgaccttgta ccatgcagcc actggcagcc tgcccttcat
     2341 cccctttggt gggccacggc ggaacaagga gatcatgtac cggatcacca cggagaagcc
     2401 ggctggggcc attgcaggtg cccagaggcg ggagaacggg cccctggagt ggagctacac
     2461 cctccccatc acctgccagc tgtcactggg gctgcagagc cagctggtgc ccatcctggc
     2521 caacatcctg gaggtggagc aggccaagtg ctggggcttc gaccagttct ttgcggagac
     2581 cagtgacatc ctgcagcgag ttgtcgtcca tgtcttctcc ctgtcccagg cagtcctgca
     2641 ccacatctat atccatgccc acaacacgat agccattttc caggaggccg tgcacaagca
     2701 gaccagtgtg gccccccgac accaggagta cctctttgag ggtcacctct gtgtcctcga
     2761 gcccagcgtc tcagcacagc acatcgccca cacgacggca agcagccccc tgaccctctt
     2821 cagcacagcc atccctaagg ggctggcctt cagggaccct gctctggacg tccccaagtt
     2881 cgtccccaaa gtggacctgc aggcggatta caacactgcc aagggcgtgt tgggcgccgg
     2941 ctaccaggcc ctgcggctgg cacgggccct gctggatggg caggagctaa tgtttcgggg
     3001 gctgcactgg gtcatggagg tgctccaggc cacatgcaga cggactctgg aagtggcaag
     3061 gacatccctc ctctacctca gcagcagcct gggaactgag aggttcagca gcgtggctgg
     3121 aacgcctgag atccaggaac tgaaggcggc tgcagaactg aggtccaggc tgcggactct
     3181 agcggaggtc ctctccagat gctcccaaaa tatcacggag acccaggaga gcctgagcag
     3241 cctgaaccgg gagctggtga agagccggga tcaggtacat gaggacagaa gcatccagca
     3301 gattcagtgc tgtttggaca agatgaactt catctacaaa cagttcaaga agtctaggat
     3361 gaggccaggg cttggctaca acgaggagca gattcacaag ctggataagg tgaatttcag
     3421 tcatttagcc aaaagactcc tgcaggtgtt ccaggaggag tgcgtgcaga agtatcaagc
     3481 gtccttagtc acacacggca agaggatgag ggtggtgcac gagaccagga accacctgcg
     3541 cctggttggc tgttctgtgg ctgcctgtaa cacagaagcc cagggggtcc aggagagtct
     3601 cagcaagctc ctggaagagc tatctcacca gctccttcag gaccgagcaa agggggctca
     3661 ggcctcgccg cctcccatag ctccttaccc cagccctaca cgaaaggacc tgcttctcca
     3721 catgcaagag ctctgcgagg ggatgaagct gctggcatct gacctcctgg acaacaaccg
     3781 catcatcgaa cggctaaata gagtcccagc acctcctgat gtcctcgagg actacaaaga
     3841 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     3901 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3961 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4021 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4081 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4141 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4201 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4261 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4321 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4381 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4441 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4501 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4561 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4621 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4741 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4801 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4861 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4921 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4981 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5041 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5101 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5161 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5221 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5281 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5341 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5401 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5461 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5521 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5581 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5641 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5701 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5761 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5821 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5881 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5941 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6001 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6061 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6121 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6181 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6241 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6301 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6361 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7021 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7081 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     7141 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7201 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7261 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7321 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7381 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7441 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7501 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7561 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7621 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7681 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7741 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7801 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7861 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7921 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7981 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8041 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8101 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8161 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8221 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8281 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
