LOCUS       V029850                 7491 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029850
VERSION     V029850
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..2171
                     /label="Mlkl(NM_001310613)"
                     /note="Mlkl(NM_001310613)"
                     /gene="Mlkl"
     CDS             2178..2243
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2267..2855
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2921..2969
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3171..3599
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3613..3942
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4009..4803
                     /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    4979..5100
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5149..5165)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5173..5189
                     /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(5197..5227)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5242..5263
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5551..6139)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6310..7170)
                     /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(7171..7275)
                     /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 taaattggga cagatcatca
      781 agttaggcca gctcatctat gaacagtgtg aaaagatgaa atactgccgg aaacaatgcc
      841 agcgtctagg aaaccgtgtg cacggcctgc tacagcctct ccagagactc caggcccaag
      901 gaaagaagaa cctgcccgat gacattactg ctgccctggg ccgttttgat gaagtcctga
      961 aggaggctaa ccagcagata gaaaagttca gcaagaagtc ccatatttgg aagtttgtga
     1021 gtgtgggcaa tgataagatc ctcttccatg aagtgaatga gaagctgaga gacgtctggg
     1081 aggagctgtt gctgctgctt caggtttatc attggaatac cgtttcagat gtcagccagc
     1141 cagcatcctg gcagcaggaa gatcgacagg atgcagagga agacggaaat gaaaatatga
     1201 aagttatcct gatgcagttg caaattagcg tggaagaaat caacaaaacc ctgaagcaat
     1261 gctcactaaa acccacacag gagatcccac aagatctcca aatcaaggag attccaaagg
     1321 aacatcttgg acctccgtgg accaaactga agacaagtaa aatgagcacc atttatagag
     1381 gagagtatca cagatctcca gttaccatca aagtattcaa caacccccag gccgaaagtg
     1441 ttggaatagt gaggttcact ttcaatgacg agatcaaaac catgaagaaa ttcgattctc
     1501 ccaacatctt gcgtatattt gggatttgca ttgatcaaac agtgaagccc cctgagttct
     1561 ccattgtcat ggagtactgt gaacttggaa ccctgaggga actgctggat agagaaaaag
     1621 acctcacaat gagtgtgcgc agcctcctag tcctgagggc agccagaggc ttatacaggc
     1681 tacaccattc ggaaacactc cacagaaaca tcagcagctc cagtttcctc gtagccggag
     1741 gctaccaagt aaagcttgca ggatttgagt taagcaaaac acagaattcc atcagccgga
     1801 cagcaaagag cactaaagca gagagatcca gttcaacgat atatgtctcc cctgagagac
     1861 tgaaaaatcc attttgcctt tatgacataa aagctgaaat atatagcttt ggaattgtac
     1921 tctgggaaat tgccactgga aagatcccat ttgaaggctg tgattctaag aagatccggg
     1981 agctggtggc tgaggacaag aagcaggaac cagtgggtca ggattgccct gagttgttgc
     2041 gggaaatcat taatgagtgt cgtgcccatg agccctccca acggccctct gtggacggta
     2101 ggagtctttc tggcagagaa cgaatcttgg agagactgtc tgcggttgaa gaatccacgg
     2161 acaagaaggt gggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2221 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2281 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2341 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2401 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2461 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2521 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2581 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2641 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2701 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2761 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2821 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     2881 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2941 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3001 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3061 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3121 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3181 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3241 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3301 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3361 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3421 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3481 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3541 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3601 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3721 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3781 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3841 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3901 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3961 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4021 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4081 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4141 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4201 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4261 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4321 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4381 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4441 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4501 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4561 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4621 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4681 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4741 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4801 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4861 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4921 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4981 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5041 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5101 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5161 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5221 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5281 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6421 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7321 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7381 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7441 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
