LOCUS       V030810                 8220 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030810
VERSION     V030810
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2900
                     /label="MTA1(NM_004689)"
                     /note="MTA1(NM_004689)"
                     /gene="MTA1"
     CDS             2907..2972
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2996..3584
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3650..3698
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3900..4328
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4342..4671
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4738..5532
                     /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    5708..5829
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5878..5894)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5902..5918
                     /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(5926..5956)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5971..5992
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6280..6868)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7039..7899)
                     /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(7900..8004)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cgccaacatg tacagggtcg
      781 gagactacgt ctactttgag aactcctcca gcaacccata cctgatccgg agaatcgagg
      841 agctcaacaa gacggccaat gggaacgtgg aggccaaagt ggtgtgcttc taccggaggc
      901 gggacatctc cagcaccctc atcgccctgg ccgacaagca cgcaaccctg tcagtctgct
      961 ataaggccgg accgggggcg gacaacggcg aggaagggga aatagaagag gaaatggaga
     1021 acccggaaat ggtggacctg cccgagaaac taaagcacca gctgcggcat cgggagctgt
     1081 tcctctcccg gcagctggag tctctgcccg ccacgcacat caggggcaag tgcagcgtca
     1141 ccctgctcaa cgagaccgag tcgctcaagt cctacctgga gcgggaggat ttcttcttct
     1201 attctctagt ctacgaccca cagcagaaga ccctgctggc agataaagga gagattcgag
     1261 taggaaaccg gtaccaggca gacatcaccg acttgttaaa agaaggcgag gaggatggcc
     1321 gagaccagtc caggttggag acccaggtgt gggaggcgca caacccactc acagacaagc
     1381 agatcgacca gttcctggtg gtggcccgct ctgtgggcac cttcgcacgg gccctggact
     1441 gcagcagctc cgtccgacag cccagcctgc acatgagcgc cgcagctgcc tcccgagaca
     1501 tcaccctgtt ccacgccatg gatactctcc acaagaacat ctacgacatc tccaaggcca
     1561 tctcggcgct ggtgccgcag ggcgggcccg tgctctgcag ggacgagatg gaggagtggt
     1621 ctgcatcaga ggccaacctt ttcgaggaag ccctggaaaa atatgggaag gatttcacgg
     1681 acattcagca agattttctc ccgtggaagt cgctgaccag catcattgag tactactaca
     1741 tgtggaagac caccgacaga tacgtgcagc agaaacgctt gaaagcagct gaagctgaga
     1801 gcaagttaaa gcaagtttat attcccaact ataacaagcc aaatccgaac caaatcagcg
     1861 tcaacaacgt caaggccggt gtggtgaacg gcacgggggc gccgggccag agccctgggg
     1921 ctggccgggc ctgcgagagc tgttacacca cacagtctta ccagtggtat tcttggggtc
     1981 cccctaacat gcagtgtcgt ctctgcgcat cttgttggac atattggaag aaatatggtg
     2041 gcttgaaaat gccaacccgg ttagatggag agaggccagg accaaaccgc agtaacatga
     2101 gtccccacgg cctcccagcc cggagcagcg ggagccccaa gtttgccatg aagaccaggc
     2161 aggctttcta tctgcacacg acgaagctga cgcggatcgc ccggcgcctg tgccgtgaga
     2221 tcctgcgccc gtggcacgct gcgcggcacc cctacctgcc catcaacagc gcggccatca
     2281 aggccgagtg cacggcgcgg ctgcccgaag cctcccagag cccgctggtg ctgaagcagg
     2341 cggtacgcaa gccgctggaa gccgtgcttc ggtatcttga gacccacccc cgccccccca
     2401 agcctgaccc cgtgaaaagc gtgtccagcg tgctcagcag cctgacgccc gccaaggtgg
     2461 cccccgtcat caacaacggc tcccccacca tcctgggcaa gcgcagctac gagcagcaca
     2521 acggggtgga cggcaacatg aagaagcgcc tcttgatgcc cagtaggggt ctggcaaacc
     2581 acggacaggc caggcacatg ggaccaagcc ggaacctcct gctcaacggg aagtcctacc
     2641 ccaccaaagt gcgcctgatc cgggggggct ccctgccccc agtcaagcgg cggcggatga
     2701 actggatcga cgccccggat gacgtgttct acatggccac agaggagacc aggaagatcc
     2761 gcaagctgct ctcatcctcg gaaaccaagc gtgctgcccg ccggccctac aagcccatcg
     2821 ccctgcgcca gagccaggcc ctgccgccgc ggccaccgcc acctgcgccc gtcaacgacg
     2881 agcccatcgt catcgaggac ggatccgact acaaagacca tgacggtgat tataaagatc
     2941 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     3001 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3061 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3121 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3181 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3241 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3301 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3361 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3421 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3481 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3541 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3601 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3661 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3721 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3781 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3841 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4021 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4081 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4201 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4261 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4321 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4441 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4501 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4561 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4621 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4681 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4741 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4801 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4861 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4921 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4981 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5041 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5101 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5161 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5221 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5281 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5341 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5401 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5461 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5521 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5581 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5641 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5701 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5761 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5821 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5881 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5941 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6181 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6721 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8041 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     8101 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     8161 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
