LOCUS       V033014                 7600 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033014
VERSION     V033014
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             706..2259
                     /label="RIPK3(NM_006871)"
                     /note="RIPK3(NM_006871)"
                     /gene="RIPK3"
     CDS             2266..2292
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2296..2322
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2326..2352
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2376..2964
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3030..3078
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3280..3708
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3722..4051
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4118..4912
                     /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    5088..5209
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5258..5274)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5282..5298
                     /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(5306..5336)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5351..5372
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5660..6248)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6419..7279)
                     /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(7280..7384)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgtc gtgcgtcaag
      721 ttatggccca gcggtgcccc cgcccccttg gtgtccatcg aggaactgga gaaccaggag
      781 ctcgtcggca aaggcgggtt cggcacagtg ttccgggcgc aacataggaa gtggggctac
      841 gatgtggcgg tcaagatcgt aaactcgaag gcgatatcca gggaggtcaa ggccatggca
      901 agtctggata acgaattcgt gctgcgccta gaaggggtta tcgagaaggt gaactgggac
      961 caagatccca agccggctct ggtgactaaa ttcatggaga acggctcctt gtcggggctg
     1021 ctgcagtccc agtgccctcg gccctggccg ctcctttgcc gcctgctgaa agaagtggtg
     1081 cttgggatgt tttacctgca cgaccagaac ccggtgctcc tgcaccggga cctcaagcca
     1141 tccaacgtcc tgctggaccc agagctgcac gtcaagctgg cagattttgg cctgtccaca
     1201 tttcagggag gctcacagtc agggacaggg tccggggagc cagggggcac cctgggctac
     1261 ttggccccag aactgtttgt taacgtaaac cggaaggcct ccacagccag tgacgtctac
     1321 agcttcggga tcctaatgtg ggcagtgctt gctggaagag aagttgagtt gccaaccgaa
     1381 ccatcactcg tgtacgaagc agtgtgcaac aggcagaacc ggccttcatt ggctgagctg
     1441 ccccaagccg ggcctgagac tcccggctta gaaggactga aggagctaat gcagctctgc
     1501 tggagcagtg agcccaagga cagaccctcc ttccaggaat gcctaccaaa aactgatgaa
     1561 gtcttccaga tggtggagaa caatatgaat gctgctgtct ccacggtaaa ggatttcctg
     1621 tctcagctca ggagcagcaa taggagattt tctatcccag agtcaggcca aggagggaca
     1681 gaaatggatg gctttaggag aaccatagaa aaccagcact ctcgtaatga tgtcatggtt
     1741 tctgagtggc taaacaaact gaatctagag gagcctccca gctctgttcc taaaaaatgc
     1801 ccgagcctta ccaagaggag cagggcacaa gaggagcagg ttccacaagc ctggacagca
     1861 ggcacatctt cagattcgat ggcccaacct ccccagactc cagagacctc aactttcaga
     1921 aaccagatgc ccagccctac ctcaactgga acaccaagtc ctggaccccg agggaatcag
     1981 ggggctgaga gacaaggcat gaactggtcc tgcaggaccc cggagccaaa tccagtaaca
     2041 gggcgaccgc tcgttaacat atacaactgc tctggggtgc aagttggaga caacaactac
     2101 ttgactatgc aacagacaac tgccttgccc acatggggct tggcaccttc gggcaagggg
     2161 aggggcttgc agcacccccc accagtaggt tcgcaagaag gccctaaaga tcctgaagcc
     2221 tggagcaggc cacagggttg gtataatcat agcgggaaac tcgagtaccc atacgacgta
     2281 cctgattatg caacttatcc ttatgatgta cctgattatg ctagctatcc ttatgatgtc
     2341 ccagactacg catagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt
     2401 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2461 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2521 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2581 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2641 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2701 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2761 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     2821 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2881 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2941 tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga
     3001 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3061 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     3121 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     3181 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     3241 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3301 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3361 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3421 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3481 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3541 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3601 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     3661 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3721 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3781 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3841 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3901 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3961 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4021 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4081 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4141 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4201 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4261 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4321 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4381 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4441 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4501 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4561 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4621 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4681 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4741 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4801 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4861 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4921 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4981 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5041 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5101 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5161 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5221 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5281 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5341 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5401 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5461 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5521 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5581 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5821 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6061 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6301 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6361 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6421 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6481 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6541 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6601 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6661 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6721 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6781 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6841 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6901 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6961 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7021 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7081 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7141 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7201 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7261 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7321 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7381 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7441 cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7501 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     7561 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
