LOCUS       V021290                 8540 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021290
VERSION     V021290
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..3187
                     /label="TRIM28(NM_005762)"
                     /note="TRIM28(NM_005762)"
                     /gene="TRIM28"
     CDS             3227..3292
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3316..3904
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3970..4018
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4220..4648
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4662..4991
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5058..5852
                     /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    6028..6149
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6198..6214)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6222..6238
                     /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(6246..6276)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6291..6312
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6600..7188)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7359..8219)
                     /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(8220..8324)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcggc ctccgcggcg gcagcctcgg cagcagcggc
      721 ctcggccgcc tctggcagcc cgggcccggg cgagggctcc gctggcggcg aaaagcgctc
      781 caccgcccct tcggccgcag cctcggcctc tgcctcagcc gcggcgtcgt cgcccgcggg
      841 gggcggcgcc gaggcgctgg agctgctgga gcactgcggc gtgtgcagag agcgcctgcg
      901 acccgagagg gagccccgcc tgctgccctg tttgcactcg gcctgtagtg cctgcttagg
      961 gcccgcggcc cccgccgccg ccaacagctc gggggacggc ggggcggcgg gcgacggcac
     1021 cgtggtggac tgtcccgtgt gcaagcaaca gtgcttctcc aaagacatcg tggagaatta
     1081 tttcatgcgt gatagtggca gcaaggctgc caccgacgcc caggatgcga accagtgctg
     1141 cactagctgt gaggataatg ccccagccac cagctactgt gtggagtgct cggagcctct
     1201 gtgtgagacc tgtgtagagg cgcaccagcg ggtgaagtac accaaggacc atactgtgcg
     1261 ctctactggg ccagccaagt ctcgggatgg tgaacgtact gtctattgca acgtacacaa
     1321 gcatgaaccc cttgtgctgt tttgtgagag ctgtgatact ctcacctgcc gagactgcca
     1381 gctcaatgcc cacaaggacc accagtacca gttcttagag gatgcagtga ggaaccagcg
     1441 caagctcctg gcctcactgg tgaagcgcct tggggacaaa catgcaacat tgcagaagag
     1501 caccaaggag gttcgcagct caatccgcca ggtgtctgac gtacagaagc gtgtgcaagt
     1561 ggatgtcaag atggccatcc tgcagatcat gaaggagctg aataagcggg gccgtgtgct
     1621 ggtcaatgat gcccagaagg tgactgaggg gcagcaggag cgcctggagc ggcagcactg
     1681 gaccatgacc aagatccaga agcaccagga gcacattctg cgctttgcct cttgggctct
     1741 ggagagtgac aacaacacag cccttttgct ttctaagaag ttgatctact tccagctgca
     1801 ccgggccctc aagatgattg tggatcccgt ggagccacat ggcgagatga agtttcagtg
     1861 ggacctcaat gcctggacca agagtgccga ggcctttggc aagattgtgg cagagcgtcc
     1921 tggcactaac tcaacaggcc ctgcacccat ggcccctcca agagccccag ggcccctgag
     1981 caagcagggc tctggcagca gccagcccat ggaggtgcag gaaggctatg gctttgggtc
     2041 aggagatgat ccctactcaa gtgcagagcc ccatgtgtca ggtgtgaaac ggtcccgctc
     2101 aggtgagggc gaggtgagcg gccttatgcg caaggtgcca cgagtgagcc ttgaacgcct
     2161 ggacctggac ctcacagctg acagccagcc acccgtcttc aaggtcttcc caggcagtac
     2221 cactgaggac tacaacctta ttgttattga acgtggcgct gccgctgcag ctaccggcca
     2281 gccagggact gcgcctgcag gaacccctgg tgccccaccc ctggctggca tggccattgt
     2341 caaggaggag gagacggagg ctgccattgg agcccctcct actgccactg agggccctga
     2401 gaccaaacct gtgcttatgg ctcttgcgga gggtcctggt gctgagggtc cccgcctggc
     2461 ctcacctagt ggcagcacca gctcagggct ggaggtggtg gctcctgagg gtacctcagc
     2521 cccaggtggt ggcccgggaa ccctggatga cagtgccacc atttgccgtg tctgccagaa
     2581 gccaggcgat ctggttatgt gcaaccagtg tgagttttgt ttccacctgg actgtcacct
     2641 gccggccctg caggatgtac caggggagga gtggagctgc tcactctgcc atgtgctccc
     2701 tgacctgaag gaggaggatg gcagcctcag cctggatggt gcagacagca ctggcgtggt
     2761 ggccaagctc tcaccagcca accagcggaa atgtgagcgt gtactgctgg ccctattctg
     2821 tcacgaaccc tgccgccccc tgcatcagct ggctaccgac tccaccttct ccctggacca
     2881 gcccggtggc accctggatc tgaccctgat ccgtgcccgc ctccaggaga agttgtcacc
     2941 tccctacagc tccccacagg agtttgccca ggatgtgggc cgcatgttca agcaattcaa
     3001 caagttaact gaggacaagg cagacgtgca gtccatcatc ggcctgcagc gcttcttcga
     3061 gacgcgcatg aacgaggcct tcggtgacac caagttctct gctgtgctgg tggagccccc
     3121 gccgatgagc ctgcctggtg ctggcctgag ttcccaggag ctgtctggtg gccctggtga
     3181 tggccccgaa ttctgcagat atccagcaca gtggcggccg ctcgaggact acaaagacca
     3241 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     3301 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3361 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3421 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3481 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3541 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3601 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3661 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     3721 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3781 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3841 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3901 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3961 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     4021 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4081 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4141 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     4201 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4261 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4321 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     4381 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4441 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4501 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4561 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     4621 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4681 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4741 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4801 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4861 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4921 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4981 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     5041 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5101 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5161 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5221 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5281 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5341 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5401 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5461 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5521 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5581 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5641 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5701 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5761 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5821 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     5881 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5941 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6001 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6061 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6121 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6181 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6241 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6301 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6361 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6421 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6481 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6541 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6601 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6661 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6721 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6781 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6841 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6901 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6961 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7021 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7081 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7141 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7201 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7261 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7321 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7381 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7441 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7501 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7561 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7621 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7681 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7741 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7801 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7861 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7921 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7981 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8041 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8101 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8161 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8221 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8281 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8341 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     8401 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     8461 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     8521 gcatgaagaa tctgcttagg
//
