LOCUS       V046409                 7800 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046409
VERSION     V046409
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..2480
                     /label="Rnf180(NM_027934)"
                     /note="Rnf180(NM_027934)"
                     /gene="Rnf180"
     CDS             2487..2552
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2576..3164
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3230..3278
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3480..3908
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3922..4251
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4318..5112
                     /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    5288..5409
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5458..5474)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5482..5498
                     /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(5506..5536)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5551..5572
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5860..6448)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6619..7479)
                     /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(7480..7584)
                     /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 ccaccatgaa aagaagcgaa gagtcgacaa
      781 gtacgcagag ccccgaggag caaacaggca cccttcattg ttggagatgt agaaaatgca
      841 ttgcaagttc tggttgcttc atgactcctc ttgagactca agtggttgag caggacagac
      901 atgaatcagt tgatgctcaa aacacctgtc acctgtggca catgaatgta gacgcccttc
      961 cggagtggat cagctgcttg ctgcagaagg cccagtggac agttgggaaa ctgaattgtc
     1021 ctttctgtgg ggcccgatta gggggcttta attttgtcag cactccaaaa tgttcctgtg
     1081 gccagcttgc agctgtacat ctctgcaaga gccggactga tcaccaagca gcacagggag
     1141 ggagactaat gagaccagcg ctgaaacatt tgccacatcc cggagtcccg tcaggttgtg
     1201 ataaggaaac tctgctgaca ggtggtggct ccaaaaccag aaaccactgg cttttaagca
     1261 tggcccgaaa cagtaacggc ctcggaagac ttacagaagc actctgcctg gaggtgcgag
     1321 cgacgtattt tgagatgaag aatgaaaaac tgctcttcaa agcctcggac ccaaaatgcc
     1381 agccttttgt tcctcagcct gacaccggca ggtgtccttc aagagcgtct cacagaaagt
     1441 cacacagttt ggacctgaac atcagtgaga agctgatttt attacccact ttatatgaaa
     1501 tacatcgtaa gcctactgcc tatcccaggc taaatgaaac ggggcccatt gacctttcag
     1561 gcttggcttt accgtgtagt aacagtagct gctcctttca aagcccacct agttttgatc
     1621 cgaatatgct gctgcacaga ctttcagtgg ctccccatga gacccaggca caaaggggaa
     1681 gagaatgcca gtgcggccta gaagcttctt cagtctactc ggatcacgct aacgctaaca
     1741 gcctgccatt cctgatggat ctgccctcag cagggaggag cgtgctggag gcctcggacc
     1801 aagaggagca cctctcccag ctggacttcc tgcgctctgc cagcttcccc ttgggcacca
     1861 ttaaccacag gctgaacaac agggagagaa gcaagttgag gactctgcga aggcagcaga
     1921 ggcgcgaacg atggctacag aagcagggta aatactcagg agtgggcttg ctggatcaca
     1981 tgaccgtgag taatgagatg agtacagatg aggaaacgga gttcccagag gagaaggaca
     2041 gctacatgtg tgcagtgtgc ctggatgtgt acttcaaccc ctacatgtgc tacccctgcc
     2101 accacatctt ctgcgagccc tgcctacgaa ctctggccaa agacaacccc gcaagcactc
     2161 cctgcccact gtgccggaca attatttcta gagtctttct ccagacagaa ctgaacaatg
     2221 ccacaaagac attctttact aaagaatatt tgaaaatcaa gcaaagcttt cagaaatcca
     2281 gctctgcgaa gtggcccttg ccgagctgca ggaaaggatt ccatctcttc ggagggaagc
     2341 cagcacaaag actcaagccg tgcaggatct tggatacagg aatgagatac gaggccatgg
     2401 aggggtgctg cttactgacc tgctcgcatg gatggcgcgc tcagcctgct ttcttatccc
     2461 actcaggaca ctgcctgggg ggatccgact acaaagacca tgacggtgat tataaagatc
     2521 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2581 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2641 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2701 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2761 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2821 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2881 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2941 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3001 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3061 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3121 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3181 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3241 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3301 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3361 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3421 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3481 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3541 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3601 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3661 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3721 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3781 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3841 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3901 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4021 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4081 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4141 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4201 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4261 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4321 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4381 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4441 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4501 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4561 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4621 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4681 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4741 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4801 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4861 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4921 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4981 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5041 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5101 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5161 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5221 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5281 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5401 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5461 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5521 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5581 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5641 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5701 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5761 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5821 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5881 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6061 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6181 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6241 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6301 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6361 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6421 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6541 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6601 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6661 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6721 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6781 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6841 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6901 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6961 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7021 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7081 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7141 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7201 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7261 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7321 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7381 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7441 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7501 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7561 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7621 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7681 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7741 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
