LOCUS       V028975                 7988 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028975
VERSION     V028975
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..2736
                     /label="Il1r1(NM_008362)"
                     /note="Il1r1(NM_008362)"
                     /gene="Il1r1"
     polyA_signal    2747..3369
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3411..3708
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3762..4556
                     /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    5208..5342
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(5348..5366)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(5380..5396)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5404..5420
                     /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(5428..5458)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5473..5494
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5782..6370)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6541..7401)
                     /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(7402..7506)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7533..7961
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgat ggagaatatg
     1021 aaagtgctac tggggctcat ttgtctcatg gtgcctctgc tgtcgctgga gattgacgta
     1081 tgtacagaat atccaaatca gatcgttttg tttttatctg taaatgaaat tgatattcgc
     1141 aagtgtcctc ttactccaaa taaaatgcac ggcgacacca taatttggta caagaatgac
     1201 agcaagaccc ccatatcagc ggaccgggac tccaggattc atcagcagaa tgaacatctt
     1261 tggtttgtac ctgccaaggt ggaggactca ggatattact attgtatagt aagaaactca
     1321 acttactgcc tcaaaactaa agtaaccgta actgtgttag agaatgaccc tggcttgtgt
     1381 tacagcacac aggccacctt cccacagcgg ctccacattg ccggggatgg aagtcttgtg
     1441 tgcccttatg tgagttattt taaagatgaa aataatgagt tacccgaggt ccagtggtat
     1501 aagaactgta aacctctgct tcttgacaac gtgagcttct tcggagtaaa agataaactg
     1561 ttggtgagga atgtggctga agagcacaga ggggactata tatgccgtat gtcctatacg
     1621 ttccggggga agcaatatcc ggtcacacga gtaatacaat ttatcacaat agatgaaaac
     1681 aagagggaca gacctgttat cctgagccct cggaatgaga cgatcgaagc tgacccagga
     1741 tcaatgatac aactgatctg caacgtcacg ggccagttct cagaccttgt ctactggaag
     1801 tggaatggat cagaaattga atggaatgat ccatttctag ctgaagacta tcaatttgtg
     1861 gaacatcctt caaccaaaag aaaatacaca ctcattacaa cacttaacat ttcagaagtt
     1921 aaaagccagt tttatcgcta tccgtttatc tgtgttgtta agaacacaaa tatttttgag
     1981 tcggcgcatg tgcagttaat atacccagtc cctgacttca agaattacct catcgggggc
     2041 tttatcatcc tcacggctac aattgtatgc tgtgtgtgca tctataaagt cttcaaggtt
     2101 gacatagtgc tttggtacag ggactcctgc tctggttttc ttccttcaaa agcttcagat
     2161 ggaaagacat acgatgccta tattctttat cccaagaccc tgggagaggg gtccttctca
     2221 gacttagata cttttgtttt taaactgttg cctgaggtct tggagggaca gtttggatac
     2281 aagctgttca tttatggaag ggatgactat gttggagaag ataccatcga ggttactaat
     2341 gaaaatgtaa agaaaagcag gaggctgatt atcattctag tgagagatat gggaggcttc
     2401 agctggctgg gccagtcatc tgaagagcaa atagccatat acaatgctct catccaggaa
     2461 ggaattaaaa tcgtcctgct tgagttggag aaaatccaag actatgagaa aatgccagat
     2521 tctattcagt tcattaagca gaaacacgga gtcatttgct ggtcaggaga ctttcaagaa
     2581 agaccacagt ctgcaaagac caggttctgg aaaaacttaa gataccagat gccagcccaa
     2641 cggagatcac cattgtctaa acaccgctta ctaaccctgg atcctgtgcg ggacactaag
     2701 gagaaactgc cggcagcaac acacttacca ctcggctagg aattccgggt ggcatccctg
     2761 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2821 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2881 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2941 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3001 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3061 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3121 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3181 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3241 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3301 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3361 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3481 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3541 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3601 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3661 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca
     3721 gcttgggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4561 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4621 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4681 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc
     4741 cctcgcgagt tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca
     4801 gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga
     4861 actgcaggag tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg
     4921 cctgatacag aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct
     4981 gaggtcactg cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc
     5041 cactcgccaa gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5101 cgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5161 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5221 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5281 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5341 tcaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     5401 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5461 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5521 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5581 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5641 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5701 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5761 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5821 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5881 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5941 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6001 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6061 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6121 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6181 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6241 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6301 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7501 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     7561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     7621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7741 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     7801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     7861 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     7921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     7981 tacaattt
//
