LOCUS       V039633                 8983 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039633
VERSION     V039633
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             964..4473
                     /label="DGKD(NM_003648)"
                     /note="DGKD(NM_003648)"
                     /gene="DGKD"
     CDS             4480..4545
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4583..4807
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4853..5281
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5295..5624
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5691..6485
                     /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    6659..6780
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6829..6845)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6853..6869
                     /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(6877..6907)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6922..6943
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7231..7816)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7987..8847)
                     /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(8848..8952)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgaata tgtttctgta ctttcagacc atcatcaaag aggggatgct gaccaaacag
     1021 aacaattcat tccagcgatc aaaaaggaga tactttaagc ttcgagggcg aacgctttac
     1081 tatgccaaaa cggcaaagtc aatcatattt gatgaggtgg atctgacaga tgccagcgta
     1141 gctgaatcca gtaccaaaaa cgtcaacaac agttttacgg tcataactcc atgcaggaag
     1201 ctcatcttgt gtgctgataa cagaaaagaa atggaagatt ggattgcagc attaaagact
     1261 gtgcagaaca gggagcactt tgagcccacc cagtacagca tggaccactt ctcagggatg
     1321 cacaattggt acgcctgttc ccacgcgagg ccgacctact gcaatgtgtg ccgtgaggct
     1381 ctgtctgggg tcacgtcgca cgggctgtcc tgcgaggtgt gcaaatttaa ggcccacaag
     1441 cgctgtgctg tgcgtgcaac caataactgc aagtggacca cactggcctc gatcgggaag
     1501 gacatcattg aagatgcaga tgggattgca atgccccacc agtggttgga aggaaaccta
     1561 cctgtgagcg ccaagtgcac tgtgtgcgac aagacctgtg gcagtgtgct gcgcctgcag
     1621 gactggcgct gcctctggtg caaggccatg gttcacacat cgtgtaaaga atccttgctg
     1681 accaagtgcc cacttggcct gtgcaaagtg tcagtcatcc cacccacggc tctcaacagc
     1741 atcgactccg atgggttctg gaaggccagc tgtcctcctt cttgcacaag cccactgttg
     1801 gtcttcgtca attcaaaaag tggggacaac cagggtgtga agttcctcag aagattcaaa
     1861 cagctactaa accccgccca ggtcttcgac ctcatgaacg gaggcccaca cctcggctta
     1921 cggttattcc agaagtttga cacattccgg attctggttt gtggcgggga tggaagtgtt
     1981 ggctgggtcc tctccgaaat cgacagcctc aaccttcata aacagtgtca gctgggagtg
     2041 ctgccgctcg gcacagggaa cgacttggcc cgagtactgg gctggggctc agcctgcgat
     2101 gacgacaccc agctccccca gatcttggag aagttggaga gagccagcac caagatgctg
     2161 gacaggtgga gcgtcatggc atacgaggcc aagctccccc ggcaggcctc ctcctctacc
     2221 gtcaccgaag acttcagcga ggattccgag gtacagcaga ttctcttcta tgaagactcg
     2281 gttgcagccc acctttctaa aatcctcacc tcggaccagc actcggtggt catctcctcg
     2341 gccaaagtgc tctgtgagac ggtgaaggac ttcgtggcac gggtggggaa ggcctatgag
     2401 aagacgaccg agagctcgga ggagtcagag gtcatggcca agaagtgctc tgtcctgaaa
     2461 gagaagctgg attcccttct caagaccttg gacgatgagt cccaggcctc gtcctctctg
     2521 cccaacccgc cccccaccat tgccgaggag gctgaagatg gagatgggtc gggcagcatc
     2581 tgcggttcca ccggagaccg cttggtggca tcagcttgcc cggcccggcc gcagatattc
     2641 cggcctcgag aacagctcat gctgagagcc aacagcctga agaaagcaat tcgtcagatc
     2701 atagaacaca cagaaaaagc tgtcgatgag cagaatgccc agacccagga gcaggagggc
     2761 ttcgtcctgg gcctctctga gtcagaggag aagatggacc acagagtgtg cccaccactg
     2821 tcccacagcg agagcttcgg ggtccccaag gggaggagcc agcgcaaagt gtcgaaatct
     2881 ccgtgtgaaa agctgatcag caaagggagt ctgtccctag gcagttctgc ttcccttccg
     2941 ccccagccgg gaagccggga cggcctgcct gcgctcaaca ccaagatcct gtacccaaat
     3001 gtccgggctg gaatgtctgg ttccttaccc ggtggctcag tcatcagtcg cctgttaatt
     3061 aatgctgatc ccttcaactc tgaaccagaa accctagagt attacacgga gaaatgtgtc
     3121 atgaacaact attttggcat tggcctggat gcgaagatat ccctggactt taacaacaag
     3181 cgcgatgagc acccagagaa gtgcaggagc cgaaccaaga acatgatgtg gtatggagtt
     3241 cttggaacca aagagttgct gcacagaacc tacaagaacc tggagcaaaa ggtcttgctg
     3301 gagtgtgacg ggcgacccat cccactcccc agtcttcagg gaattgctgt ccttaacatt
     3361 cccagctatg ccggaggaac caacttctgg gggggtacca aggaagatga tactttcgca
     3421 gctccatcat tcgatgacaa gattctggag gtggtcgccg tgttcggcag catgcagatg
     3481 gccgtctctc gagtcatcag gctacagcat catcggatcg cccagtgtcg cacggtgaag
     3541 atctccatcc ttggggatga gggcgtgcct gtgcaggtgg acggagaggc ctgggtccag
     3601 ccgccagggt acattcggat tgtccacaag aaccgggcac agacactgac cagagacagg
     3661 gcatttgaga gcaccctgaa gtcctgggaa gacaagcaga agtgcgagct gccccgccct
     3721 ccatcctgtt ccctgcaccc ggagatgctg tccgaggagg aggccaccca gatggaccag
     3781 tttgggcagg cagcaggggt cctcattcac agtatccgag aaatagctca gtctcaccgg
     3841 gacatggagc aggaactggc ccacgccgtc aatgccagct ccaagtccat ggaccgtgtg
     3901 tatggcaagc ccagaaccac agaggggctc aactgcagct tcgtcctgga aatggtgaat
     3961 aacttcagag ctctgcgcag tgagacggag ctgctgctgt ctgggaagat ggccctgcag
     4021 ctggatccgc ctcagaagga gcagctgggg agtgctcttg ccgagatgga ccgacagctc
     4081 aggaggctgg cagacacccc gtggctctgc cagtccgcag agcccggcga cgaagagagt
     4141 gtgatgctgg atcttgccaa gcgcagtcgc agtggtaaat tccgcctcgt gaccaagttt
     4201 aaaaaggaga aaaacaacaa gaacaaagaa gctcacagta gcctgggagc cccggttcac
     4261 ctctggggga cagaggaggt tgctgcctgg ctggagcacc tcagtctctg tgagtataag
     4321 gacatcttca cacggcacga catccggggc tctgagctcc tgcacctgga gcggagggac
     4381 ctcaaggacc tgggcgtgac caaggtgggc cacatgaaga ggatcctgtg tggcatcaag
     4441 gagctgagcc gcagcgcccc cgccgtcgag gccctcgagg actacaaaga ccatgacggt
     4501 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgtt
     4561 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4621 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4681 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4741 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4801 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4861 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4921 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4981 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5041 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5101 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5161 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5221 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5281 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5341 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5401 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5461 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5521 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5581 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5641 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5701 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5761 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5821 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5881 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5941 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6001 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6061 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6121 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6181 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     6241 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6301 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6361 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6421 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6481 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6541 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6601 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6661 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6721 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6781 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6841 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6901 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6961 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7021 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7081 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7741 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     7801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
