LOCUS       V040994                 6810 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040994
VERSION     V040994
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..1490
                     /label="Ywhaz(NM_011740)"
                     /note="Ywhaz(NM_011740)"
                     /gene="Ywhaz"
     CDS             1497..1562
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1586..2174
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2240..2288
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2490..2918
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2932..3261
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3328..4122
                     /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    4298..4419
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4468..4484)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4492..4508
                     /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(4516..4546)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4561..4582
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4870..5458)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5629..6489)
                     /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(6490..6594)
                     /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 ccaccatgga taaaaatgag ctggtgcaga
      781 aggccaagct ggccgagcag gcagagcgat atgatgacat ggcagcctgc atgaagtctg
      841 tcactgagca gggagctgag ctgtcgaatg aggagagaaa ccttctctct gttgcttata
      901 aaaacgttgt aggagcccgt aggtcatcgt ggagggtcgt ctcaagtatt gagcagaaga
      961 cggaaggtgc tgagaaaaag cagcagatgg ctcgagaata cagagagaag atcgagacgg
     1021 agctgcgtga catctgcaac gatgtactgt ctcttttgga aaagttcttg atccccaatg
     1081 cttcgcaacc agaaagcaaa gtcttctatt tgaaaatgaa gggtgactac taccgttact
     1141 tggccgaggt tgctgctggt gatgacaaga aaggaattgt ggaccagtca cagcaagcat
     1201 accaagaagc atttgaaatc agcaaaaagg agatgcagcc gacacacccc atcagactgg
     1261 gtctggccct caacttctct gtgttctatt acgagatcct gaactcccca gagaaagcct
     1321 gctctcttgc aaaaacagct ttcgatgaag ccattgctga acttgataca ttaagtgaag
     1381 agtcgtacaa agacagcacg ctaataatgc agttactgag agacaactta acattgtgga
     1441 catcggatac ccaaggagat gaagcagaag caggagaagg aggggaaaat ggatccgact
     1501 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     1561 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     1621 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1681 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1741 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     1801 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     1861 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     1921 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     1981 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2041 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2101 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2161 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     2221 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     2281 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     2341 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     2401 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     2461 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2521 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2581 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2641 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2701 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2761 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2821 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     2881 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2941 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3001 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3061 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3121 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3181 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3241 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3301 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3361 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3421 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3481 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3541 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3601 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3661 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3721 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3781 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3841 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3901 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3961 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4021 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4081 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4141 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4201 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4261 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4321 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4381 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4441 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4501 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4561 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4621 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4681 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5041 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5281 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6601 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     6661 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     6721 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     6781 accgacaatt gcatgaagaa tctgcttagg
//
