LOCUS       V033683                 6490 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033683
VERSION     V033683
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             919..2241
                     /label="EED(NM_003797)"
                     /note="EED(NM_003797)"
                     /gene="EED"
     CDS             2254..2319
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2354..2578
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2624..3052
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3066..3395
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3407..4006
                     /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    4166..4287
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4336..4352)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4360..4376
                     /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(4384..4414)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4429..4450
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4738..5323)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5494..6354)
                     /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(6355..6459)
                     /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 gagccaccat gtccgagagg gaagtgtcga ctgcgccggc gggaacagac
      961 atgcctgcgg ccaagaagca gaagctgagc agtgacgaga acagcaatcc agacctctct
     1021 ggagacgaga atgatgacgc tgtcagtata gaaagtggta caaacactga acgccctgat
     1081 acacctacaa acacgccaaa tgcacctgga aggaaaagtt ggggaaaggg aaaatggaag
     1141 tcaaagaaat gcaaatattc tttcaaatgt gtaaatagtc tcaaggaaga tcataaccaa
     1201 ccattgtttg gagttcagtt taactggcac agtaaagaag gagatccatt agtgtttgca
     1261 actgtaggaa gcaacagagt taccttgtat gaatgtcatt cacaaggaga aatccggttg
     1321 ttgcaatctt acgtggatgc tgatgctgat gaaaactttt acacttgtgc atggacctat
     1381 gatagcaata cgagccatcc tctgctggct gtagctggat ctagaggcat aattaggata
     1441 ataaatccta taacaatgca gtgtataaag cactatgttg gccatggaaa tgctatcaat
     1501 gagctgaaat tccatccaag agatccaaat cttctcctgt cagtaagtaa agatcatgct
     1561 ttacgattat ggaatatcca gacggacact ctggtggcaa tatttggagg cgtagaaggg
     1621 cacagagatg aagttctaag tgctgattat gatcttttgg gtgaaaaaat aatgtcctgt
     1681 ggtatggatc attctcttaa actttggagg atcaattcaa agagaatgat gaatgcaatt
     1741 aaggaatctt atgattataa tccaaataaa actaacaggc catttatttc tcagaaaatc
     1801 cattttcctg atttttctac cagagacata cataggaatt atgttgattg tgtgcgatgg
     1861 ttaggcgatt tgatactttc taagtcttgt gaaaatgcca ttgtgtgctg gaaacctggc
     1921 aagatggaag atgatataga taaaattaaa cccagtgaat ctaatgtgac tattcttggg
     1981 cgatttgatt acagccagtg tgacatttgg tacatgaggt tttctatgga tttctggcaa
     2041 aagatgcttg cattgggcaa tcaagttggc aaactttatg tttgggattt agaagtagaa
     2101 gatcctcata aagccaaatg tacaacactg actcatcata aatgtggtgc tgctattcga
     2161 caaaccagtt ttagcaggga tagcagcatt cttatagctg tttgtgatga tgccagtatt
     2221 tggcgctggg atcgacttcg actcgagacc ggtgactaca aagaccatga cggtgattat
     2281 aaagatcatg acatcgatta caaggatgac gatgacaagt gaccggtaag tttaaaccgc
     2341 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2401 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2461 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2521 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2581 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2641 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2701 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2761 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2821 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2881 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2941 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggccatt
     3001 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3061 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3121 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3241 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3301 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3361 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggatga ccgagtacaa
     3421 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3481 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3541 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3601 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3661 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3721 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3781 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3841 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3901 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3961 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgattcg aaatgaccga
     4021 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4081 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4141 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4261 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4501 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     5281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6481 acctgacgtc
//
