LOCUS       V036530                 7336 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036530
VERSION     V036530
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             937..3288
                     /label="RIPK4(NM_020639)"
                     /note="RIPK4(NM_020639)"
                     /gene="RIPK4"
     CDS             3301..3366
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3401..3625
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3671..4099
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4113..4442
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4454..4852
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     polyA_signal    5012..5133
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5182..5198)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5206..5222
                     /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(5230..5260)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5275..5296
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5584..6169)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6340..7200)
                     /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(7201..7305)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcga tatcggatcc gccaccatgg agggcgacgg cgggacccca
      961 tgggccctgg cgctgctgcg caccttcgac gcgggcgagt tcacgggctg ggagaaggtg
     1021 ggctcgggcg gcttcgggca ggtgtacaag gtgcgccatg tccactggaa gacctggctg
     1081 gccatcaagt gctcgcccag cctgcacgtc gacgacaggg agcgcatgga gcttttggaa
     1141 gaagccaaga agatggagat ggccaagttt cgctacatcc tgcctgtgta tggcatctgc
     1201 cgcgaacctg tcggcctggt catggagtac atggagacgg gctccctgga aaagctgctg
     1261 gcttcggagc cattgccatg ggatctccgg ttccgaatca tccacgagac ggcggtgggc
     1321 atgaacttcc tgcactgcat ggccccgcca ctcctgcacc tggacctcaa gcccgcgaac
     1381 atcctgctgg atgcccacta ccacgtcaag atttctgatt ttggtctggc caagtgcaac
     1441 gggctgtccc actcgcatga cctcagcatg gatggcctgt ttggcacaat cgcctacctc
     1501 cctccagagc gcatcaggga gaagagccgg ctcttcgaca ccaagcacga tgtatacagc
     1561 tttgcgatcg tcatctgggg cgtgctcaca cagaagaagc cgtttgcaga tgagaagaac
     1621 atcctgcaca tcatggtgaa ggtggtgaag ggccaccgcc ccgagctgcc gcccgtgtgc
     1681 agagcccggc cgcgcgcctg cagccacctg atacgcctca tgcagcggtg ctggcagggg
     1741 gatccgcgag ttaggcccac cttccaagaa attacttctg aaaccgagga cctgtgtgaa
     1801 aagcctgatg acgaagtgaa agaaactgct catgatctgg acgtgaaaag ccccccggag
     1861 cccaggagcg aggtggtgcc tgcgaggctc aagcgggcct ctgcccccac cttcgataac
     1921 gactacagcc tctccgagct gctctcacag ctggactctg gagtttccca ggctgtcgag
     1981 ggccccgagg agctcagccg cagctcctct gagtccaagc tgccatcgtc cggcagtggg
     2041 aagaggctct cgggggtgtc ctcggtggac tccgccttct cttccagagg atcactgtcg
     2101 ctgtcctttg agcgggaacc ttcaaccagc gatctgggca ccacagacgt ccagaagaag
     2161 aagcttgtgg atgccatcgt gtccggggac accagcaaac tgatgaagat cctgcagccg
     2221 caggacgtgg acctggcact ggacagcggt gccagcctgc tgcacctggc ggtggaggcc
     2281 gggcaagagg agtgcgccaa gtggctgctg ctcaacaatg ccaaccccaa cctgagcaac
     2341 cgtaggggct ccaccccgtt gcacatggcc gtggagagga gggtgcgggg tgtcgtggag
     2401 ctcctgctgg cgcggaagat cagtgtcaac gccaaggatg aggaccagtg gacagccctc
     2461 cactttgcag cccagaacgg ggacgagtct agcacacggc tgctgttgga gaagaacgcc
     2521 tcggtcaacg aggtggactt tgagggccgg acgcccatgc acgtggcctg ccagcacggg
     2581 caggagaata tcgtgcgcat cctgctgcgc cgaggcgtgg acgtgagcct gcagggcaag
     2641 gatgcctggc tgccactgca ctacgctgcc tggcagggcc acctgcccat cgtcaagctg
     2701 ctggccaagc agccgggggt gagtgtgaac gcccagacgc tggatgggag gacgccattg
     2761 cacctggccg cacagcgcgg gcactaccgc gtggcccgca tcctcatcga cctgtgctcc
     2821 gacgtcaacg tctgcagcct gctggcacag acacccctgc acgtggccgc ggagacgggg
     2881 cacacgagca ctgccaggct gctcctgcat cggggcgctg gcaaggaggc catgacctca
     2941 gacggctaca ccgctctgca cctggctgcc cgcaacggac acctggccac tgtcaagctg
     3001 cttgtcgagg agaaggccga tgtgctggcc cggggacccc tgaaccagac ggcgctgcac
     3061 ctggctgccg cccacgggca ctcggaggtg gtggaggagt tggtcagcgc cgatgtcatt
     3121 gacctgttcg acgagcaggg gctcagcgcg ctgcacctgg ccgcccaggg ccggcacgca
     3181 cagacggtgg agactctgct caggcatggg gcccacatca acctgcagag cctcaagttc
     3241 cagggcggcc atggccccgc cgccacgctc ctgcggcgaa gcaagaccct cgagaccggt
     3301 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     3361 gacaagtgac cggtaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     3421 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3481 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3541 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3601 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3661 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3721 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3781 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3841 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3901 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3961 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4021 tttgatttat aagggatttt ggccatttcg gcctattggt taaaaaatga gctgatttaa
     4081 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4201 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4261 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4321 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4381 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4441 aaaagctccc gggatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     4501 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     4561 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     4621 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     4681 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     4741 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     4801 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aattcgaaat
     4861 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4921 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4981 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5161 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7321 agtgccacct gacgtc
//
