LOCUS       V023872                 6389 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023872
VERSION     V023872
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             941..1879
                     /label="RRAGA(NM_006570)"
                     /note="RRAGA(NM_006570)"
                     /gene="RRAGA"
     CDS             1886..1951
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1989..2213
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2259..2687
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2701..3030
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3097..3891
                     /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    4065..4186
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4235..4251)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4259..4275
                     /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(4283..4313)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4328..4349
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4637..5222)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5393..6253)
                     /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(6254..6358)
                     /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 atccgccacc atgccaaata cagccatgaa
      961 gaaaaaggtg ctgctgatgg ggaagagcgg gtcggggaag accagcatga ggtcgataat
     1021 cttcgccaat tacattgctc gcgacacccg gcgcctgggg gccaccattg acgtggaaca
     1081 ctcccacgtc cgattcctag ggaacctggt gctgaacctg tgggactgtg gcggtcagga
     1141 caccttcatg gaaaattact tcaccagcca gcgagacaat atcttccgta acgtggaagt
     1201 tttgatttac gtgtttgacg tggagagccg cgaactggaa aaggacatgc attattacca
     1261 gtcgtgtctg gaggccatcc tccagaactc tcctgacgcc aaaatcttct gcctggtgca
     1321 caaaatggat ctggttcagg aggatcagcg tgacctgatt tttaaagagc gagaggaaga
     1381 cctgaggcgt ctgtctcgcc cgctggagtg tgcttgtttt cgaacgtcca tctgggatga
     1441 gacgctctac aaagcctggt ccagcatcgt ctaccagctg attcccaacg ttcagcagct
     1501 ggagatgaac ctcaggaatt ttgcccaaat cattgaggcc gatgaagttc tgctgttcga
     1561 aagagctaca ttcttggtta tttcccacta ccagtgcaaa gagcagcgcg acgtccaccg
     1621 gtttgagaag atcagcaaca tcatcaaaca gttcaagctg agctgcagta aattggccgc
     1681 ttccttccag agcatggaag ttaggaattc caacttcgct gctttcatcg acatcttcac
     1741 ctcaaatacg tacgtgatgg tggtcatgtc agatccgtcg atcccttctg cggccactct
     1801 gatcaacatt cgcaatgccc ggaaacactt tgagaagctg gagagagtgg atggccccaa
     1861 gcacagtctc cttatgcgtc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1921 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     1981 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2041 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2101 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2161 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2221 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2281 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2341 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2401 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2461 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2521 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2581 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2641 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2701 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2761 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2821 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2881 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2941 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3001 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3061 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3121 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3181 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3241 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3301 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3361 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3421 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3481 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3541 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3601 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3661 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3721 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3781 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3841 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3901 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3961 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4021 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4201 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4261 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4321 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4381 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4441 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5041 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5161 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5341 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5401 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5461 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5521 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5581 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5641 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5701 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5761 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5821 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5881 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5941 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6001 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6061 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6121 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6181 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6241 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6301 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6361 catttccccg aaaagtgcca cctgacgtc
//
