LOCUS       .                       9523 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    205..793
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             865..1098
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1176..1310
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1337..1472
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1493..1511)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1521..1537)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      1679..2134
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2160..2264
                     /label="AmpR promoter"
     CDS             2265..3122
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3296..3884
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4172..4193
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4208..4238
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4246..4262
                     /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)."
     primer_bind     4270..4286
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4307..4325
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4353..4579
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4580..4760
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4807..4932
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5425..5658
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5843..5887
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     promoter        6062..6562
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             6574..7596
                     /label="HygR"
                     /note="hygromycin B phosphotransferase"
     misc_feature    7650..7767
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        7884..8187
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8188..8391
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             8402..9163
                     /label="miniTurbo"
                     /note="N-terminally truncated mutant of the E. coli biotin
                     ligase BioID engineered for improved catalytic efficiency
                     (Branon et al., 2018)"
     CDS             9230..9505
                     /label="FRB"
                     /note="FKBP-rapamycin binding domain of human FRAP"
ORIGIN
        1 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtcctcgagc tcaagcttcg
       61 aattcgagtg ctggtggtat caccaccgtg gagtccaact cctcctggtg gaccaactgg
      121 gtgatccccg ccatctccgc cctggtggtg gccctgatgt accgcctgta catggccgag
      181 gactagtaaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
      241 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
      301 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
      361 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
      421 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
      481 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
      541 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
      601 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
      661 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
      721 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
      781 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
      841 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
      901 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      961 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     1021 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     1081 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     1141 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     1201 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1261 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     1321 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     1441 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     1501 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1681 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1741 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1801 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1861 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1921 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1981 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2041 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2101 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3901 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3961 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4021 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4081 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4141 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4201 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4261 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     4321 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     4381 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     4441 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     4501 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     4561 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     4621 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4681 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4741 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     4801 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     4861 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     4921 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     4981 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     5041 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     5101 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     5161 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     5221 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     5281 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     5341 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     5401 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     5461 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     5521 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     5581 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     5641 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     5701 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     5761 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     5821 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     5881 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     5941 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     6001 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     6061 cttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     6121 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     6181 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     6241 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     6301 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     6361 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     6421 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     6481 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     6541 aatcaccgac ctctctcccc agggggtacc accatgaaaa agcctgaact caccgctacc
     6601 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcc
     6661 gagggcgaag aatctcgggc tttcagcttc gatgtgggag ggcgtggata tgtcctgcgg
     6721 gtgaatagct gcgccgatgg tttctacaaa gatcgctatg tttatcggca ctttgcatcc
     6781 gccgctctcc ctattcccga agtgcttgac attggggagt tcagcgagag cctgacctat
     6841 tgcatctccc gccgtgcaca gggtgtcacc ttgcaagacc tgcctgaaac cgaactgccc
     6901 gctgttctcc agcccgtcgc cgaggccatg gatgccatcg ctgccgccga tcttagccag
     6961 accagcgggt tcggcccatt cggacctcaa ggaatcggtc aatacactac atggcgcgat
     7021 ttcatctgcg ctattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     7081 gtcagtgcct ccgtcgccca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     7141 gaagtccggc acctcgtgca cgccgatttc ggctccaaca atgtcctgac cgacaatggc
     7201 cgcataacag ccgtcattga ctggagcgag gccatgttcg gggattccca atacgaggtc
     7261 gccaacatct tcttctggag gccctggttg gcttgtatgg agcagcagac ccgctacttc
     7321 gagcggaggc atcccgagct tgcaggatct cctcggctcc gggcttatat gctccgcatt
     7381 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggct
     7441 cagggtcgct gcgacgcaat cgtccggtcc ggagccggga ctgtcgggcg tacacaaatc
     7501 gcccgcagaa gcgctgccgt ctggaccgat ggctgtgtgg aagtgctcgc cgatagtgga
     7561 aacagacgcc ccagcactcg tcctagggca aaggatctgc agtaatgaga attctagatc
     7621 ttgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     7681 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     7741 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt
     7801 tggcgccggc tcgagggggc ccggggttat taatagtaat caattacggg gtcattagtt
     7861 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     7921 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     7981 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     8041 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     8101 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     8161 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     8221 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     8281 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     8341 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     8401 actgtcggga tccgccacca tgggacagct ggacggcggg agcgtggcag tcctgcctgt
     8461 ggtcgactcc accaatcagt acctgctgga tcgaatcggc gagctgaaga gtggggatgc
     8521 ttgcattgca gaatatcagc aggcagggag aggaagcaga gggaggaaat ggttctctcc
     8581 ttttggagct aacctgtacc tgagtatgtt ttggcgcctg aagcggggac cagcagcaat
     8641 cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg ctgcgaaagc tgggagcaga
     8701 caaggtgcga gtcaaatggc ccaatgacct gtatctgcag gatagaaagc tggcaggcat
     8761 cctggtggag ctggccggaa taacaggcga tgctgcacag atcgtcattg gcgccgggat
     8821 taacgtggct atgaggcgcg tggaggaaag cgtggtcaat cagggctgga tcacactgca
     8881 ggaagcaggg attaacctgg acaggaatac tctggccgct acgctgatcc gagagctgcg
     8941 ggcagccctg gaactgttcg agcaggaagg cctggctcca tatctgccac ggtgggagaa
     9001 gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag agattttcgg
     9061 gattagccgg gggattgata aacagggagc cctgctgctg gaacaggacg gagttatcaa
     9121 accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aagggctcgg gcgggagtac
     9181 cggttaccca tacgacgttc cagactacgg tagtgggagc ggtgtggcca tcctgtggca
     9241 cgagatgtgg catgaaggcc tggaagaggc atctcgtttg tactttgggg aaaggaacgt
     9301 gaaaggcatg tttgaggtgc tggagccctt gcatgctatg atggaacggg gcccccagac
     9361 tctgaaggaa acatccttta atcaggccta tggtcgagat ttaatggagg cccaagagtg
     9421 gtgcaggaag tacatgaaat cagggaatgt caaggacctc acccaagcct gggacctcta
     9481 ttatcatgtg ttccgacgaa tctcaagtgc tggtggtagt gct
//
