LOCUS       .                       8786 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        582..785
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        830..848
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3380..3400
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             3419..4132
                     /label="mEGFP"
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     CDS             4166..4189
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin®, an engineered form
                     of streptavidin"
     CDS             4226..4249
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin®, an engineered form
                     of streptavidin"
     CDS             4283..4309
                     /label="9xHis"
                     /note="9xHis affinity tag"
     polyA_signal    4353..4577
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4623..5051
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5065..5394
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5461..6252
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6429..6562
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6599..6615)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6623..6639)
                     /label="lac operator"
                     /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)."
     promoter        complement(6647..6677)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6692..6713)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7001..7586)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7760..8617)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(8618..8722)
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gagagcgagc agctgttcca ccgcggctac taccgcaaca gctacaacag
      961 catcaccagc gccagcagcg acgaggagct gctggacggc gccggcgcca tcatggactt
     1021 ccagaccagc gaggacgaca acctgctgga cggcgacacc gccgccggca cccactacac
     1081 catgaccaac ggcggcagca tcaacagcag cacccacctg ctggacctgc tggacgagcc
     1141 catccccggc gtgggcacct acgacgactt ccacaccatc gactgggtgc gcgagaagtg
     1201 caaggaccgc gagcgccacc gccgcatcaa cagcaagaag aaggagagcg cctgggagat
     1261 gaccaagagc ctgtacgacg cctggagcgg ctggctggtg gtgaccctga ccggcctggc
     1321 cagcggcgcc ctggccggcc tgatcgacat cgccgccgac tggatgaccg acctgaagga
     1381 gggcatctgc ctgagcgccc tgtggtacaa ccacgagcag tgctgctggg gcagcaacga
     1441 gaccaccttc gaggagcgcg acaagtgccc ccagtggaag acctgggccg agctgatcat
     1501 cggccaggcc gagggccccg gcagctacat catgaactac atcatgtaca tcttctgggc
     1561 cctgagcttc gccttcctgg ccgtgagcct ggtgaaggtg ttcgccccct acgcctgcgg
     1621 cagcggcatc cccgagatca agaccatcct gagcggcttc atcatccgcg gctacctggg
     1681 caagtggacc ctgatgatca agaccatcac cctggtgctg gccgtggcca gcggcctgag
     1741 cctgggcaag gagggccccc tggtgcacgt ggcctgctgc tgcggcaaca tcttcagcta
     1801 cctgttcccc aagtacagca ccaacgaggc caagaagcgc gaggtgctga gcgccgccag
     1861 cgccgccggc gtgagcgtgg ccttcggcgc ccccatcggc ggcgtgctgt tcagcctgga
     1921 ggaggtgagc tactacttcc ccctgaagac cctgtggcgc agcttcttcg ccgccctggt
     1981 ggccgccttc gtgctgcgca gcatcaaccc cttcggcaac agccgcctgg tgctgttcta
     2041 cgtggagtac cacaccccct ggtacctgtt cgagctgttc cccttcatcc tgctgggcgt
     2101 gttcggcggc ctgtggggcg ccttcttcat ccgcgccaac atcgcctggt gccgccgccg
     2161 caagagcacc aagttcggca agtaccccgt gctggaggtg atcatcgtgg ccgccatcac
     2221 cgccgtgatc gccttcccca acccctacac ccgcctgaac accagcgagc tgatcaagga
     2281 gctgttcacc gactgcggcc ccctggagag cagcagcctg tgcgactacc gcaacgacat
     2341 gaacgccagc aagatcgtgg acgacatccc cgaccgcccc gccggcgtgg gcgtgtacag
     2401 cgccatctgg cagctgtgcc tggccctgat cttcaagatc atcatgaccg tgttcacctt
     2461 cggcatcaag gtgcccagcg gcctgttcat ccccagcatg gccatcggcg ccatcgccgg
     2521 ccgcatcgtg ggcatcgccg tggagcagct ggcctactac caccacgact ggttcatctt
     2581 caaggagtgg tgcgaggtgg gcgccgactg catcaccccc ggcctgtacg ccatggtggg
     2641 cgccgccgcc tgcctgggcg gcgtgacccg catgaccgtg agcctggtgg tgatcgtgtt
     2701 cgagctgacc ggcggcctgg agtacatcgt gcccctgatg gccgccgtga tgaccagcaa
     2761 gtgggtgggc gacgccttcg gccgcgaggg catctacgag gcccacatcc gcctgaacgg
     2821 ctaccccttc ctggacgcca aggaggagtt cacccacacc accctggccg ccgacgtgat
     2881 gcgcccccgc cgcagcgacc cccccctggc cgtgctgacc caggacaaca tgaccgtgga
     2941 cgacatcgag aacatgatca acgagaccag ctacaacggc ttccccgtga tcatgagcaa
     3001 ggagagccag cgcctggtgg gcttcgccct gcgccgcgac ctgaccatcg ccatcgagag
     3061 cgcccgcaag aagcaggagg gcatcgtggg cagcagccgc gtgtgcttcg cccagcacac
     3121 ccccagcctg cccgccgaga gcccccgccc cctgaagctg cgcagcatcc tggacatgag
     3181 ccccttcacc gtgaccgacc acacccccat ggagatcgtg gtggacatct tccgcaagct
     3241 gggcctgcgc cagtgcctgg tgacccacaa cggcatcgtg ctgggcatca tcaccaagaa
     3301 ggacatcctg cgccacatgg cccagaccgc caaccaggac cccgccagca tcatgttcaa
     3361 cctcgagttc gaatctagag aaaaccttta cttccagggc aacggcagca gcggatctgt
     3421 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3481 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3541 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3601 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3661 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3721 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3781 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3841 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3901 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3961 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     4021 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     4081 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggtggagg
     4141 ttcgggaggt ggatcgggtg gaagctggag ccacccgcag ttcgagaaag gtggaggttc
     4201 gggaggtgga tcgggtggaa gcgcttggag ccacccgcag ttcgagaaag gtggaggttc
     4261 gggaggtgga tcgggtggaa gccaccatca ccatcaccat caccatcacc attaatctag
     4321 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4381 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4441 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4501 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4561 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4621 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4681 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4741 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4801 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4861 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4921 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4981 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5041 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5101 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5161 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5221 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5281 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5341 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5401 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5461 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5521 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5581 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5641 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5701 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5761 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5821 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5881 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5941 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6001 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6061 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6121 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6181 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6241 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6301 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6361 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6421 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6541 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7441 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7501 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     7561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7801 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7861 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7921 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7981 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8041 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8101 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8161 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8221 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8281 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8341 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8401 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8461 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8521 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8581 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8641 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8701 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8761 cgggagatct cccgatcccc tatggt
//
