LOCUS       V033527                 6529 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033527
VERSION     V033527
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..2280
                     /label="FBLN5(NM_006329)"
                     /note="FBLN5(NM_006329)"
                     /gene="FBLN5"
     CDS             2293..2358
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2393..2617
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2663..3091
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3105..3434
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3446..4045
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4205..4326
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4375..4391)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4399..4415
                     /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(4423..4453)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4468..4489
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4777..5362)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5533..6393)
                     /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(6394..6498)
                     /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 gccaccatgc caggaataaa aaggatactc
      961 actgttacca ttctggctct ctgtcttcca agccctggga atgcacaggc acagtgcacg
     1021 aatggctttg acctggatcg ccagtcagga cagtgtttag atattgatga atgccgaacc
     1081 atccccgagg cctgccgagg agacatgatg tgtgttaacc aaaatggcgg gtatttatgc
     1141 attccccgga caaaccctgt gtatcgaggg ccctactcga acccctactc gaccccctac
     1201 tcaggtccgt acccagcagc tgccccacca ctctcagctc caaactatcc cacgatctcc
     1261 aggcctctta tatgccgctt tggataccag atggatgaaa gcaaccaatg tgtggatgtg
     1321 gacgagtgtg caacagattc ccaccagtgc aaccccaccc agatctgcat caatactgaa
     1381 ggcgggtaca cctgctcctg caccgacgga tattggcttc tggaaggcca gtgcttagac
     1441 attgatgaat gtcgctatgg ttactgccag cagctctgtg cgaatgttcc tggatcctat
     1501 tcttgtacat gcaaccctgg ttttaccctc aatgaggatg gaaggtcttg ccaagatgtg
     1561 aacgagtgtg ccaccgagaa cccctgcgtg caaacctgcg tcaacaccta cggctctttc
     1621 atctgccgct gtgacccagg atatgaactt gaggaagatg gcgttcattg cagtgatatg
     1681 gacgagtgca gcttctctga gttcctctgc caacatgagt gtgtgaacca gcccggcaca
     1741 tacttctgct cctgccctcc aggctacatc ctgctggatg acaaccgaag ctgccaagac
     1801 atcaacgaat gtgagcacag gaaccacacg tgcaacctgc agcagacgtg ctacaattta
     1861 caagggggct tcaaatgcat tgaccccatc cgctgtgagg agccttatct gaggatcagt
     1921 gataaccgct gtatgtgtcc tgctgagaac cctggctgca gagaccagcc ctttaccatc
     1981 ttgtaccggg acatggacgt ggtgtcagga cgctccgttc ccgctgacat cttccaaatg
     2041 caagccacga cccgctaccc tggggcctat tacattttcc agatcaaatc tgggaatgag
     2101 ggcagagaat tttacatgcg gcaaacgggc cccatcagtg ccaccctggt gatgacacgc
     2161 cccatcaaag ggccccggga aatccagctg gacttggaaa tgatcactgt caacactgtc
     2221 atcaacttca gaggcagctc cgtgatccga ctgcggatat atgtgtcgca gtacccattc
     2281 ctcgagaccg gtgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     2341 aaggatgacg atgacaagtg accggtaagt ttaaaccgct gatcagcctc gactgtgcct
     2401 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2461 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2521 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2581 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2641 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2701 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2761 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2821 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2881 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2941 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3001 tcggtctatt cttttgattt ataagggatt ttggccattt cggcctattg gttaaaaaat
     3061 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3181 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3241 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3301 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3361 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3421 taggcttttg caaaaagctc ccgggatgac cgagtacaag cccacggtgc gcctcgccac
     3481 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3541 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3601 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3661 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3721 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3781 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3841 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     3901 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3961 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4021 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc
     4081 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4141 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4201 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4261 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4321 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4381 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4441 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4501 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4561 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4621 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4681 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4741 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5281 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     5341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
