LOCUS       V022511                 8666 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022511
VERSION     V022511
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             941..4123
                     /label="NPR1(NM_000906)"
                     /note="NPR1(NM_000906)"
                     /gene="NPR1"
     CDS             4163..4228
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4266..4490
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4536..4964
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4978..5307
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5374..6168
                     /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    6342..6463
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6512..6528)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6536..6552
                     /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(6560..6590)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6605..6626
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6914..7499)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7670..8530)
                     /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(8531..8635)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgccggggc cccggcgccc
      961 cgctggctcc cgcctgcgcc tgctcctgct cctgctgctg ccgccgctgc tgctgctgct
     1021 ccggggcagc cacgcgggca acctgacggt agccgtggta ctgccgctgg ccaatacctc
     1081 gtacccctgg tcgtgggcgc gcgtgggacc cgccgtggag ctggccctgg cccaggtgaa
     1141 ggcgcgcccc gacttgctgc cgggctggac ggtccgcacg gtgctgggca gcagcgaaaa
     1201 cgcgctgggc gtctgctccg acaccgcagc gcccctggcc gcggtggacc tcaagtggga
     1261 gcacaacccc gctgtgttcc tgggccccgg ctgcgtgtac gccgccgccc cagtggggcg
     1321 cttcaccgcg cactggcggg tcccgctgct gaccgccggc gccccggcgc tgggcttcgg
     1381 tgtcaaggac gagtatgcgc tgaccacccg cgcggggccc agctacgcca agctggggga
     1441 cttcgtggcg gcgctgcacc gacggctggg ctgggagcgc caagcgctca tgctctacgc
     1501 ctaccggccg ggtgacgaag agcactgctt cttcctcgtg gaggggctgt tcatgcgggt
     1561 ccgcgaccgc ctcaatatta cggtggacca cctggagttc gccgaggacg acctcagcca
     1621 ctacaccagg ctgctgcgga ccatgccgcg caaaggccga gttatctaca tctgcagctc
     1681 ccctgatgcc ttcagaaccc tcatgctcct ggccctggaa gctggcttgt gtggggagga
     1741 ctacgttttc ttccacctgg atatctttgg gcaaagcctg caaggtggac agggccctgc
     1801 tccccgcagg ccctgggaga gaggggatgg gcaggatgtc agtgcccgcc aggcctttca
     1861 ggctgccaaa atcattacat ataaagaccc agataatccc gagtacttgg aattcctgaa
     1921 gcagttaaaa cacctggcct atgagcagtt caacttcacc atggaggatg gcctggtgaa
     1981 caccatccca gcatccttcc acgacgggct cctgctctat atccaggcag tgacggagac
     2041 tctggcacat gggggaactg ttactgatgg ggagaacatc actcagcgga tgtggaaccg
     2101 aagctttcaa ggtgtgacag gatacctgaa aattgatagc agtggcgatc gggaaacaga
     2161 cttctccctc tgggatatgg atcccgagaa tggtgccttc agggttgtac tgaactacaa
     2221 tgggacttcc caagagctgg tggctgtgtc ggggcgcaaa ctgaactggc ccctggggta
     2281 ccctcctcct gacatcccca aatgtggctt tgacaacgaa gacccagcat gcaaccaaga
     2341 tcacctttcc accctggagg tgctggcttt ggtgggcagc ctctccttgc tcggcattct
     2401 gattgtctcc ttcttcatat acaggaagat gcagctggag aaggaactgg cctcggagct
     2461 gtggcgggtg cgctgggagg acgttgagcc cagtagcctt gagaggcacc tgcggagtgc
     2521 aggcagccgg ctgaccctga gcgggagagg ctccaattac ggctccctgc taaccacaga
     2581 gggccagttc caagtctttg ccaagacagc atattataag ggcaacctcg tggctgtgaa
     2641 acgtgtgaac cgtaaacgca ttgagctgac acgaaaagtc ctgtttgaac tgaagcatat
     2701 gcgggatgtg cagaatgaac acctgaccag gtttgtggga gcctgcaccg acccccccaa
     2761 tatctgcatc ctcacagagt actgtccccg tgggagcctg caggacattc tggagaatga
     2821 gagcatcacc ctggactgga tgttccggta ctcactcacc aatgacatcg tcaagggcat
     2881 gctgtttcta cacaatgggg ctatctgttc ccatgggaac ctcaagtcat ccaactgcgt
     2941 ggtagatggg cgctttgtgc tcaagatcac cgactatggg ctggagagct tcagggacct
     3001 ggacccagag caaggacaca ccgtttatgc caaaaagctg tggacggccc ctgagctcct
     3061 gcgaatggct tcaccccctg tgcggggctc ccaggctggt gacgtataca gctttgggat
     3121 catccttcag gagattgccc tgaggagtgg ggtcttccac gtggaaggtt tggacctgag
     3181 ccccaaagag atcatcgagc gggtgactcg gggtgagcag ccccccttcc ggccctccct
     3241 ggccctgcag agtcacctgg aggagttggg gctgctcatg cagcggtgct gggctgagga
     3301 cccacaggag aggccaccat tccagcagat ccgcctgacg ttgcgcaaat ttaacaggga
     3361 gaacagcagc aacatcctgg acaacctgct gtcccgcatg gagcagtacg cgaacaatct
     3421 ggaggaactg gtggaggagc ggacccaggc atacctggag gagaagcgca aggctgaggc
     3481 cctgctctac cagatcctgc ctcactcagt ggctgagcag ctgaagcgtg gggagacggt
     3541 gcaggccgaa gcctttgaca gtgttaccat ctacttcagt gacattgtgg gtttcacagc
     3601 gctgtcggcg gagagcacac ccatgcaggt ggtgaccctg ctcaatgacc tgtacacttg
     3661 ctttgatgct gtcatagaca actttgatgt gtacaaggtg gagacaattg gcgatgccta
     3721 catggtggtg tcagggctcc ctgtgcggaa cgggcggcta cacgcctgcg aggtagcccg
     3781 catggccctg gcactgctgg atgctgtgcg ctccttccga atccgccacc ggccccagga
     3841 gcagctgcgc ttgcgcattg gcatccacac aggacctgtg tgtgctggag tggtgggact
     3901 gaagatgccc cgttactgtc tctttgggga tacagtcaac acagcctcaa gaatggagtc
     3961 taatggggaa gccctgaaga tccacttgtc ttctgagacc aaggctgtcc tggaggagtt
     4021 tggtggtttc gagctggagc ttcgagggga tgtagaaatg aagggcaaag gcaaggttcg
     4081 gacctactgg ctccttgggg agagggggag tagcacccga ggcgaattct gcagatatcc
     4141 agcacagtgg cggccgctcg aggactacaa agaccatgac ggtgattata aagatcatga
     4201 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     4261 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4321 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4381 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4441 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4501 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4561 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4621 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4681 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4741 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4801 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4861 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4921 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4981 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5041 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5101 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5161 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5221 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5281 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5341 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5401 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5461 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5521 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5581 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5641 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5701 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5761 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5821 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5881 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5941 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6001 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6061 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6121 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6181 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6241 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6301 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6361 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6421 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6481 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7321 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7441 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7501 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7561 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7621 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7681 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7741 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7801 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7861 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7921 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7981 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8041 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8101 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8161 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8221 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8281 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8341 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8401 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8461 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8521 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8581 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8641 ttccccgaaa agtgccacct gacgtc
//
