LOCUS       V034949                 6499 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034949
VERSION     V034949
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             925..2250
                     /label="CRBN(NM_016302)"
                     /note="CRBN(NM_016302)"
                     /gene="CRBN"
     CDS             2263..2328
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2363..2587
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2633..3061
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3075..3404
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3416..4015
                     /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    4175..4296
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4345..4361)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4369..4385
                     /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(4393..4423)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4438..4459
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4747..5332)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5503..6363)
                     /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(6364..6468)
                     /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 gagaattcgc caccatggcc ggcgaaggag atcagcagga cgctgcgcac
      961 aacatgggca accacctgcc gctcctgcct gcagagagtg aggaagaaga tgaaatggaa
     1021 gttgaagacc aggatagtaa agaagccaaa aaaccaaaca tcataaattt tgacaccagt
     1081 ctgccgacat cacatacata cctaggtgct gatatggaag aatttcatgg caggactttg
     1141 cacgatgacg acagctgtca ggtgattcca gttcttccac aagtgatgat gatcctgatt
     1201 cccggacaga cattacctct tcagcttttt caccctcaag aagtcagtat ggtgcggaat
     1261 ttaattcaga aagatagaac ctttgctgtt cttgcataca gcaatgtaca ggaaagggaa
     1321 gcacagtttg gaacaacagc agagatatat gcctatcgag aagaacagga ttttggaatt
     1381 gagatagtga aagtgaaagc aattggaaga caaaggttca aagtccttga gctaagaaca
     1441 cagtcagatg gaatccagca agctaaagtg caaattcttc ccgaatgtgt gttgccttca
     1501 accatgtctg cagttcaatt agaatccctc aataagtgcc agatatttcc ttcaaaacct
     1561 gtctcaagag aagaccaatg ttcatataaa tggtggcaga aataccagaa gagaaagttt
     1621 cattgtgcaa atctaacttc atggcctcgc tggctgtatt ccttatatga tgctgagacc
     1681 ttaatggaca gaatcaagaa acagctacgt gaatgggatg aaaatctaaa agatgattct
     1741 cttccttcaa atccaataga tttttcttac agagtagctg cttgtcttcc tattgatgat
     1801 gtattgagaa ttcagctcct taaaattggc agtgctatcc agcgacttcg ctgtgaatta
     1861 gacattatga ataaatgtac ttccctttgc tgtaaacaat gtcaagaaac agaaataaca
     1921 accaaaaatg aaatattcag tttatcctta tgtgggccga tggcagctta tgtgaatcct
     1981 catggatatg tgcatgagac acttactgtg tataaggctt gcaacttgaa tctgataggc
     2041 cggccttcta cagaacacag ctggtttcct gggtatgcct ggactgttgc ccagtgtaag
     2101 atctgtgcaa gccatattgg atggaagttt acggccacca aaaaagacat gtcacctcaa
     2161 aaattttggg gcttaacgcg atctgctctg ttgcccacga tcccagacac tgaagatgaa
     2221 ataagtccag acaaagtaat actttgcttg ctcgagaccg gtgactacaa agaccatgac
     2281 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagtg accggtaagt
     2341 ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2401 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2461 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2521 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2581 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2641 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2701 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2761 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2821 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2881 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2941 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3001 ttggccattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3061 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggatgac
     3421 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3481 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3541 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3601 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3661 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3721 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3781 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3841 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3901 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3961 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcga
     4021 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4081 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4141 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4201 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4261 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4321 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4381 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4441 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4501 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5281 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6481 aaaagtgcca cctgacgtc
//
