LOCUS       .                       8780 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             3374..3394
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             3413..4126
                     /label="mEGFP"
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     CDS             4160..4183
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin®, an engineered form
                     of streptavidin"
     CDS             4220..4243
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin®, an engineered form
                     of streptavidin"
     CDS             4277..4303
                     /label="9xHis"
                     /note="9xHis affinity tag"
     polyA_signal    4347..4571
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4617..5045
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5059..5388
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5455..6246
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6423..6556
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6593..6609)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6617..6633)
                     /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(6641..6671)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6686..6707)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6995..7580)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7754..8611)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(8612..8716)
                     /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 gccatgtggc agggcgccat ggacaaccgc ggcttccacc agggcagctt
      961 cagcagcttc cagagcagca gcagcgacga ggacctgatg gacatccccg gcaccgccat
     1021 ggacttcagc atgcgcgacg acgtgccccc cctggaccgc gagatcgagg gcaacaagag
     1081 ctacaacggc ggcggcatcg gcagcagcaa ccgcgtgatg gacttcctgg aggagcccat
     1141 ccccggcgtg ggcacctacg acgacttcaa caccatcgac tgggtgcgcg agaagagccg
     1201 cgaccgcgac cgccaccgcg agatcaccaa caagagcaag gagagcacct gggccctgat
     1261 ccacagcgtg agcgacgcct tcagcggctg gctgctgatg ctgctgatcg gcctgctgag
     1321 cggcagcctg gccggcctga tcgacatcag cgcccactgg atgaccgacc tgaaggaggg
     1381 catctgcacc ggcggcttct ggttcaacca cgagcactgc tgctggaaca gcgagcacgt
     1441 gaccttcgag caccgcgaca agtgccccga gtggaacagc tgggcccagc tgatcatcaa
     1501 caccgaccag ggcgccttcg cctacatcgt gaactacttc atgtacgtgc tgtgggccct
     1561 gctgttcgcc ttcctggccg tgagcctggt gaaggccttc gccccctacg cctgcggcag
     1621 cggcatcccc gagatcaaga ccatcctgag cggcttcatc atccgcggct acctgggcaa
     1681 gtggaccctg gtgatcaaga ccatcaccct ggtgctggcc gtgagcagcg gcctgagcct
     1741 gggcaaggag ggccccctgg tgcacgtggc ctgctgctgc ggcaacatcc tgtgccactg
     1801 cttcaacaag taccgcaaga acgaggccaa gcgccgcgag gtgctgagcg ccgccgccgc
     1861 cgccggcgtg agcgtggcct tcggcgcccc catcggcggc gtgctgttca gcctggagga
     1921 ggtgagctac tacttccccc tgaagaccct gtggcgcagc ttcttcgccg ccctggtggc
     1981 cgccttcacc ctgcgcagca tcaacccctt cggcaacagc cgcctggtgc tgttctacgt
     2041 ggagttccac accccctggc acctgttcga gctggtgccc ttcatcgtgc tgggcatctt
     2101 cggcggcctg tggggcgccc tgttcatccg caccaacatc gcctggtgcc gcaagcgcaa
     2161 gaccacccag ctgggcaagt accccgtggt ggaggtgctg atcgtgaccg ccatcaccgc
     2221 catcctggcc ttccccaacg agtacacccg catgagcacc agcgagctga tcagcgagct
     2281 gttcaacgac tgcggcctgc tggacagcag caagctgtgc gactacgaga accacttcaa
     2341 caccagcaag ggcggcgagc tgcccgaccg ccccgccggc gtgggcatct acagcgccat
     2401 gtggcagctg gccctgaccc tgatcctgaa gatcgtgatc accatcttca ccttcggcat
     2461 gaagatcccc agcggcctgt tcatccccag catggccgtg ggcgccatcg ccggccgcct
     2521 gctgggcgtg ggcatggagc agctggccta ctaccaccac gactggggca tcttcaacag
     2581 ctggtgcagc cagggcgccg actgcatcac ccccggcctg tacgccatgg tgggcgccgc
     2641 cgcctgcctg ggcggcgtga cccgcatgac cgtgagcctg gtggtgatca tgttcgagct
     2701 gaccggcggc ctggagtaca tcgtgcccct gatggccgcc gccatgacca gcaagtgggt
     2761 ggccgacgcc ctgggccgcg agggcatcta cgacgcccac atccgcctga acggctaccc
     2821 cttcctggag gccaaggagg agttcgccca caagaccctg gccatggacg tgatgaagcc
     2881 ccgccgcaac gaccccctgc tgaccgtgct gacccaggac agcatgaccg tggaggacgt
     2941 ggagaccatc atcagcgaga ccacctacag cggcttcccc gtggtggtga gccgcgagag
     3001 ccagcgcctg gtgggcttcg tgctgcgccg cgacctgatc atcagcatcg agaacgcccg
     3061 caagaagcag gacggcgtgg tgagcaccag catcatctac ttcaccgagc acagcccccc
     3121 catgcccccc tacacccccc ccaccctgaa gctgcgcaac atcctggacc tgagcccctt
     3181 caccgtgacc gacctgaccc ccatggagat cgtggtggac atcttccgca agctgggcct
     3241 gcgccagtgc ctggtgaccc acaacggccg cctgctgggc atcatcacca agaaggacgt
     3301 gctgaagcac atcgcccaga tggccaacca ggaccccgac agcatcctgt tcaacctcga
     3361 gttcgaatct agagaaaacc tttacttcca gggcaacggc agcagcggat ctgtgagcaa
     3421 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3481 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3541 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3601 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3661 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3721 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3781 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3841 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3901 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3961 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4021 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4081 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtg gaggttcggg
     4141 aggtggatcg ggtggaagct ggagccaccc gcagttcgag aaaggtggag gttcgggagg
     4201 tggatcgggt ggaagcgctt ggagccaccc gcagttcgag aaaggtggag gttcgggagg
     4261 tggatcgggt ggaagccacc atcaccatca ccatcaccat caccattaat ctagagggcc
     4321 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4381 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4441 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4501 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4561 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     4621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4801 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4921 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4981 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5041 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5401 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5461 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5521 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5581 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5641 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5701 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5761 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5821 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5881 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5941 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6001 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6061 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6121 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6181 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6241 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7501 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     7561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7621 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7681 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7741 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7801 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7861 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7921 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7981 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8041 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8101 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8161 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8221 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8281 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8341 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8401 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8461 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8521 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8581 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8641 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8701 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8761 atctcccgat cccctatggt
//
