LOCUS       .                       6183 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             908..973
                     /label="3xFLAG"
                     /note="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
     CDS             977..1672
                     /label="BioID2"
                     /note="promiscuous R40G mutant of a biotin protein ligase
                     from Aquifex aeolicus (Kim et al., 2016)"
     polyA_signal    1779..2003
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2049..2477
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2491..2821
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2888..3679
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3856..3989
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4026..4042)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4050..4066)
                     /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(4074..4104)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4119..4140)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4428..5016)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5190..6047)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(6048..6152)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
      961 tgacgatgac aaggacttca agaacctgat ctggctgaag gaggtggaca gcacccagga
     1021 gagactgaag gagtggaacg tgagctacgg caccgccctg gtggccgaca gacagaccaa
     1081 gggcagaggc ggcctgggca gaaagtggct gagccaggag ggcggcctgt acttcagctt
     1141 cctgctgaac cccaaggagt tcgagaacct gctgcagctg cccctggtgc tgggcctgag
     1201 cgtgagcgag gccctggagg agatcaccga gatccccttc agcctgaagt ggcccaacga
     1261 cgtgtacttc caggagaaga aggtgagcgg cgtgctgtgc gagctgagca aggacaagct
     1321 gatcgtgggc atcggcatca acgtgaacca gagagagatc cccgaggaga tcaaggacag
     1381 agccaccacc ctgtacgaga tcaccggcaa ggactgggac agaaaggagg tgctgctgaa
     1441 ggtgctgaag agaatcagcg agaacctgaa gaagttcaag gagaagagct tcaaggagtt
     1501 caagggcaag atcgagagca agatgctgta cctgggcgag gaggtgaagc tgctgggcga
     1561 gggcaagatc accggcaagc tggtgggcct gagcgagaag ggcggcgccc tgatcctgac
     1621 cgaggagggc atcaaggaga tcctgagcgg cgagttcagc ctgagaagaa gcctcgagcg
     1681 gccgccactg tgctggatat ctgcagaatt ccaccacact ggactagtgg atccgagctc
     1741 ggtaccaagc ttaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc
     1801 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1861 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1921 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1981 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2041 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2101 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2161 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     2221 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2281 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2341 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2401 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     2461 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2521 ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2641 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2701 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2761 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2821 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     2881 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2941 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3001 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3061 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3121 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3181 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3241 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3301 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3361 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3421 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3481 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3541 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3601 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3661 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3721 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3781 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3841 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3901 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3961 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4021 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4081 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4141 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4201 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4261 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4321 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4381 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4441 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4501 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4561 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4621 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4681 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4741 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4801 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4861 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4921 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4981 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5041 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5101 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5161 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5221 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5281 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5341 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5401 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5461 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5521 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5581 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5641 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5701 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5761 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5821 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5881 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5941 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6001 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6061 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6121 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6181 gtc
//
