LOCUS       .                       8573 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             3167..3187
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             3206..3919
                     /label="mEGFP"
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     CDS             3953..3976
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin®, an engineered form
                     of streptavidin"
     CDS             4013..4036
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin®, an engineered form
                     of streptavidin"
     CDS             4070..4096
                     /label="9xHis"
                     /note="9xHis affinity tag"
     polyA_signal    4140..4364
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4410..4838
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4852..5181
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5248..6039
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6216..6349
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6386..6402)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6410..6426)
                     /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(6434..6464)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6479..6500)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6788..7373)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7547..8404)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(8405..8509)
                     /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 gacttcctgg aggagccctt ccccgacgtg ggcacctacg aggacttcca
      961 caccatcgac tggctgcgcg agaagagccg cgacaccgac cgccaccgca agatcaccag
     1021 caagagcaag gagagcatct gggagttcat caagagcctg ctggacgcct ggagcggctg
     1081 ggtggtgatg ctgctgatcg gcctgctggc cggcaccctg gccggcgtga tcgacctggc
     1141 cgtggactgg atgaccgacc tgaaggaggg cgtgtgcctg agcgccttct ggtacagcca
     1201 cgagcagtgc tgctggacca gcaacgagac caccttcgag gaccgcgaca agtgccccct
     1261 gtggcagaag tggagcgagc tgctgctgag ccagagcgag ggcgccagcg cctacatcct
     1321 gaactacctg atgtacatcc tgtgggccct gctgttcgcc ttcctggccg tgagcctggt
     1381 gcgcgtgttc gccccctacg cctgcggcag cggcatcccc gagatcaaga ccatcctgag
     1441 cggcttcatc atccgcggct acctgggcaa gtggaccctg ctgatcaaga ccgtgaccct
     1501 ggtgctggtg gtgagcagcg gcctgagcct gggcaaggag ggccccctgg tgcacgtggc
     1561 ctgctgctgc ggcaacttct tcagcagcct gttcagcaag tacagcaaga acgagggcaa
     1621 gcgccgcgag gtgctgagcg ccgccgccgc cgccggcgtg agcgtggcct tcggcgcccc
     1681 catcggcggc gtgctgttca gcctggagga ggtgagctac tacttccccc tgaagaccct
     1741 gtggcgcagc ttcttcgccg ccctggtggc cgccttcacc ctgcgcagca tcaacccctt
     1801 cggcaacagc cgcctggtgc tgttctacgt ggagtaccac accccctggt acatggccga
     1861 gctgttcccc ttcatcctgc tgggcgtgtt cggcggcctg tggggcaccc tgttcacccg
     1921 ctgcaacatc gcctggtgcc gccgccgcaa gaccacccgc ctgggccgct accccgtgct
     1981 ggaggtgatc gccgtgaccg ccgtgaccgc catcgtggcc taccccaacc cctacacccg
     2041 ccagagcacc agcgagctga tcagcgagct gttcaacgac tgcggcgccc tggagagcag
     2101 ccagctgtgc gactacatca acgaccccaa catgacccgc cccgtggacg acatccccga
     2161 ccgccccgcc ggcgtgggcg tgtacaccgc catgtggcag ctggccctgg ccctgatctt
     2221 caagatcgtg atcaccatct tcaccttcgg catgaagatc cccagcggcc tgttcatccc
     2281 cagcatggcc gtgggcgcca tggccggccg catggtgggc atcggcgtgg agcagctggc
     2341 ctaccaccac cacgactgga tcatcttccg caactggtgc cgccccggcg ccgactgcgt
     2401 gacccccggc ctgtacgcca tggtgggcgc cgccgcctgc ctgggcggcg tgacccgcat
     2461 gaccgtgagc ctggtggtga tcatgttcga gctgaccggc ggcctggagt acatcgtgcc
     2521 cctgatggcc gccgccgtga ccagcaagtg ggtggccgac gccttcggca aggagggcat
     2581 ctacgaggcc cacatccacc tgaacggcta ccccttcctg gacgtgaagg acgagttcac
     2641 ccaccgcacc ctggccaccg acgtgatgcg cccccgccgc ggcgagcccc ccctgagcgt
     2701 gctgacccag gacagcatga ccgtggagga cgtggagacc ctgatcaagg agaccgacta
     2761 caacggcttc cccgtgctgg tgagccgcga cagcgagcgc ctgatcggct tcgcccagcg
     2821 ccgcgagctg atcctggcca tcaagaacgc ccgccagcgc caggagggca tcgtgagcaa
     2881 cagcatcatg tacttcaccg aggagccccc cgagctgccc gccaacagcc cccaccccct
     2941 gaagctgcgc cgcatcctga acctgagccc cttcaccgtg accgaccaca cccccatgga
     3001 gaccgtggtg gacatcttcc gcaagctggg cctgcgccag tgcctggtga cccgcagcgg
     3061 ccgcctgctg ggcatcatca ccaagaagga cgtgctgcgc cacatggccc agatggccaa
     3121 ccaggacccc gagagcatca tgttcaacct cgagttcgaa tctagagaaa acctttactt
     3181 ccagggcaac ggcagcagcg gatctgtgag caagggcgag gagctgttca ccggggtggt
     3241 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3301 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3361 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3421 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3481 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3541 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3601 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3661 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3721 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3781 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     3841 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3901 catggacgag ctgtacaagg gtggaggttc gggaggtgga tcgggtggaa gctggagcca
     3961 cccgcagttc gagaaaggtg gaggttcggg aggtggatcg ggtggaagcg cttggagcca
     4021 cccgcagttc gagaaaggtg gaggttcggg aggtggatcg ggtggaagcc accatcacca
     4081 tcaccatcac catcaccatt aatctagagg gcccgtttaa acccgctgat cagcctcgac
     4141 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4201 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4261 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4321 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4381 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4441 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4501 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4561 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4621 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4681 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4741 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4801 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5101 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5161 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5221 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5281 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5341 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5401 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5461 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5521 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5581 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5641 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5701 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5761 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5821 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5881 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5941 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6001 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6061 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     6121 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6181 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6241 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6301 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6361 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6601 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7321 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7381 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7441 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7501 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7561 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7621 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7681 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7741 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7801 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7861 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7921 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7981 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8041 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8101 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8161 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8221 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8281 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8341 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8401 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8461 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8521 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
