LOCUS       .                       8756 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..1639
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     CDS             4295..4318
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     polyA_signal    4356..4580
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4626..5054
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5068..5397
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5464..6255
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6432..6565
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6602..6618)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6626..6642)
                     /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(6650..6680)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6695..6716)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7004..7589)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7763..8620)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(8621..8725)
                     /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 aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatgac
     1681 tcggcgtagg gtgctaagcg tggtcgtgct gctagccgcc ctggcgtgcc gtctcggtgc
     1741 gcagacccca gagcagcccg caccccccgc caccacggtg cagcctaccg ccacgcgtca
     1801 gcaaaccagc tttcctttcc gagtctgcga gctctccagc cacggcgacc tgttccgctt
     1861 ctcctcggac atccagtgtc cctcgtttgg cacgcgggag aatcacacgg agggcctgtt
     1921 gatggtgttt aaagacaaca ttattcccta ctcgtttaag gtccgctcct acaccaagat
     1981 agtgaccaac attctcatct acaatggctg gtacgcggac tccgtgacca accggcacga
     2041 ggagaagttc tccgttgaca gctacgaaac tgaccagatg gataccatct accagtgcta
     2101 caacgcggtc aagatgacaa aagatgggct gacgcgcgtg tatgtagacc gcgacggagt
     2161 taacatcacc gtcaacctaa agcccaccgg gggcctggcc aacggggtgc gccgctacgc
     2221 cagccagacg gagctctatg acgcccccgg gtggttgata tggacttaca gaacaagaac
     2281 taccgtcaac tgcctgataa ctgacatgat ggccaagtcc aacagcccct tcgacttctt
     2341 tgtgaccacc accgggcaga ctgtggaaat gtcccctttc tatgacggga aaaataagga
     2401 aaccttccat gagcgggcag actccttcca cgtgagaact aactacaaga tagtggacta
     2461 cgacaaccga gggacgaacc cgcaaggcga acgccgagcc ttcctggaca agggcactta
     2521 cacgctatct tggaagctcg agaacaggac agcctactgc ccgcttcaac actggcaaac
     2581 ctttgactcg accatcgcca cagaaacagg gaagtcaata cattttgtga ctgacgaggg
     2641 cacctctagc ttcgtgacca acacaaccgt gggcatagag ctcccggacg ccttcaagtg
     2701 catcgaagag caggtgaaca agaccatgca tgagaagtac gaggccgtcc aggatcgtta
     2761 cacgaagggc caggaagcca ttacatattt tataacgagc ggaggattgt tattagcttg
     2821 gctacctctg accccgcgct cgttggccac cgtcaagaac ctgacggagc ttaccactcc
     2881 gacttcctca ccccccagca gtccatcgcc cccagcccca tccgcggccc gcgggagcac
     2941 ccccgccgcc gttctgaggc gtcggaggcg ggatgcgggg aacgccacca caccggtgcc
     3001 ccccacggcc cccgggaagt ccctgggcac cctcaacaat cccgccaccg tccagatcca
     3061 atttgcctac gactccctgc gccgccagat caaccgcatg ctgggagacc ttgcgcgggc
     3121 ctggtgcctg gagcagaaga ggcagaacat ggtgctgaga gaactaacca agattaatcc
     3181 aaccaccgtc atgtccagca tctacggtaa ggcggtggcg gccaagcgcc tgggggatgt
     3241 catctcagtc tcccagtgcg tgcccgttaa ccaggccacc gtcaccctgc gcaagagcat
     3301 gagggtccct ggctccgaga ccatgtgcta ctcgcgcccc ctggtgtcct tcagctttat
     3361 caacgacacc aagacctacg agggacagct gggcaccgac aacgagatct tcctcacaaa
     3421 aaagatgacg gaggtgtgcc aggcgaccag ccagtactac ttccagtccg gcaacgagat
     3481 ccacgtctac aacgactacc accactttaa aaccatcgag ctggacggca ttgccaccct
     3541 gcagaccttc atctcactaa acacctccct catcgagaac attgactttg cctccctgga
     3601 gctgtactca cgggacgaac agcgtgcctc caacgtcttt gacctggagg gcatcttccg
     3661 ggagtacaac ttccaggcgc aaaacatcgc cggcctgcgg aaggatttgg acaatgcagt
     3721 gtcaaacgga agaaatcaat tcgtggacgg cctgggggaa cttatggaca gtctgggtag
     3781 cgtgggtcag tccatcacca acctagtcag cacggtgggg ggtttgttta gcagcctggt
     3841 ctctggtttc atctccttct tcaaaaaccc cttcggcggc atgctcattc tggtcctggt
     3901 ggcgggggtg gtgatcctgg ttatttccct cacgaggcgc acgcgccaga tgtcgcagca
     3961 gccggtgcag atgctctacc ccgggatcga cgagctcgct cagcaacatg cctctggtga
     4021 gggtccaggc attaatccca ttagtaagac agaattacaa gccatcatgt tagcgctgca
     4081 tgagcaaaac caggagcaaa agagagcagc tcagagggcg gccggaccct cagtggccag
     4141 cagagcattg caggcagcca gggaccgttt tccaggccta cgcagaagac gctatcacga
     4201 tccagagacc gccgccgcac tgcttgggga ggcagagact gagtttctcg aggactacaa
     4261 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     4321 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4381 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4441 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4501 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4561 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4621 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4681 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4741 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4801 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4861 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4921 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4981 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5041 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5101 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5161 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5221 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5281 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5341 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5401 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5461 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5521 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5581 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5641 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5701 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5761 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5821 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5881 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5941 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6001 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6061 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6121 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6181 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6241 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6301 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6361 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6421 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6481 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6541 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6601 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6661 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6721 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7441 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     7561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
