LOCUS       V022289                 6181 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022289
VERSION     V022289
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        27..330
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        331..534
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     intron          672..768
                     /note="modified SV40 intron with splice donor and acceptor
                     sites"
                     /label="SV40 intron"
     CDS             832..897
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             925..3294
                     /label="TNFAIP3(NM_001270508)"
                     /note="TNFAIP3(NM_001270508)"
                     /gene="TNFAIP3"
     polyA_signal    3320..3454
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3565..3581)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3589..3605
                     /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(3613..3643)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3658..3679
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3967..4555)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4726..5586)
                     /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(5587..5691)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     6165..6181
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggactacaaa
      841 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagctt
      901 atggccatgg aggcccgaat tcggatggct gaacaagtcc ttcctcaggc tttgtatttg
      961 agcaatatgc ggaaagctgt gaagatacgg gagagaactc cagaagacat ttttaaacct
     1021 actaatggga tcattcatca ttttaaaacc atgcaccgat acacactgga aatgttcaga
     1081 acttgccagt tttgtcctca gtttcgggag atcatccaca aagccctcat cgacagaaac
     1141 atccaggcca ccctggaaag ccagaagaaa ctcaactggt gtcgagaagt ccggaagctt
     1201 gtggcgctga aaacgaacgg tgacggcaat tgcctcatgc atgccacttc tcagtacatg
     1261 tggggcgttc aggacacaga cttggtactg aggaaggcgc tgttcagcac gctcaaggaa
     1321 acagacacac gcaactttaa attccgctgg caactggagt ctctcaaatc tcaggaattt
     1381 gttgaaacgg ggctttgcta tgatactcgg aactggaatg atgaatggga caatcttatc
     1441 aaaatggctt ccacagacac acccatggcc cgaagtggac ttcagtacaa ctcactggaa
     1501 gaaatacaca tatttgtcct ttgcaacatc ctcagaaggc caatcattgt catttcagac
     1561 aaaatgctaa gaagtttgga atcaggttcc aatttcgccc ctttgaaagt gggtggaatt
     1621 tacttgcctc tccactggcc tgcccaggaa tgctacagat accccattgt tctcggctat
     1681 gacagccatc attttgtacc cttggtgacc ctgaaggaca gtgggcctga aatccgagct
     1741 gttccacttg ttaacagaga ccggggaaga tttgaagact taaaagttca ctttttgaca
     1801 gatcctgaaa atgagatgaa ggagaagctc ttaaaagagt acttaatggt gatagaaatc
     1861 cccgtccaag gctgggacca tggcacaact catctcatca atgccgcaaa gttggatgaa
     1921 gctaacttac caaaagaaat caatctggta gatgattact ttgaacttgt tcagcatgag
     1981 tacaagaaat ggcaggaaaa cagcgagcag gggaggagag aggggcacgc ccagaatccc
     2041 atggaacctt ccgtgcccca gctttctctc atggatgtaa aatgtgaaac gcccaactgc
     2101 cccttcttca tgtctgtgaa cacccagcct ttatgccatg agtgctcaga gaggcggcaa
     2161 aagaatcaaa acaaactccc aaagctgaac tccaagccgg gccctgaggg gctccctggc
     2221 atggcgctcg gggcctctcg gggagaagcc tatgagccct tggcgtggaa ccctgaggag
     2281 tccactgggg ggcctcattc ggccccaccg acagcaccca gcccttttct gttcagtgag
     2341 accactgcca tgaagtgcag gagccccggc tgccccttca cactgaatgt gcagcacaac
     2401 ggattttgtg aacgttgcca caacgcccgg caacttcacg ccagccacgc cccagaccac
     2461 acaaggcact tggatcccgg gaagtgccaa gcctgcctcc aggatgttac caggacattt
     2521 aatgggatct gcagtacttg cttcaaaagg actacagcag aggcctcctc cagcctcagc
     2581 accagcctcc ctccttcctg tcaccagcgt tccaagtcag atccctcgcg gctcgtccgg
     2641 agcccctccc cgcattcttg ccacagagct ggaaacgacg cccctgctgg ctgcctgtct
     2701 caagctgcac ggactcctgg ggacaggacg gggacgagca agtgcagaaa agccggctgc
     2761 gtgtattttg ggactccaga aaacaagggc tttgccacac tggctttcat cgagtacaga
     2821 gaaaacaaac attttgctgc tgcctcaggg aaagtcagtc ccacagcgtc caggttccag
     2881 aacaccattc cgtgcctggg gagggaatgc ggcacccttg gaagcaccat gtttgaagga
     2941 tactgccaga agtgtttcat tgaagctcag aatcagagat ttcatgaggc caaaaggaca
     3001 gaagagcaac tgagatcgag ccagcgcaga gatgtgcctc gaaccacaca aagcacctca
     3061 aggcccaagt gcgcccgggc ctcctgcaag aacatcctgg cctgccgcag cgaggagctc
     3121 tgcatggagt gtcagcatcc caaccagagg atgggccctg gggcccaccg gggtgagcct
     3181 gcccccgaag acccccccaa gcagcgttgc cgggcccccg cctgtgatca ttttggcaat
     3241 gccaagtgca acggctactg caacgaatgc tttcagttca agcagatgta tggctaactc
     3301 gaggtaccgc ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac
     3361 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3421 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3481 gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag
     3541 gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3601 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3661 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3721 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3781 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3841 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4321 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4381 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4441 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4501 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5701 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5761 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5821 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5881 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5941 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6001 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6061 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6121 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6181 t
//
