LOCUS       .                      10081 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    20..608
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             680..913
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    991..1125
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1152..1287
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1308..1326)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1494..1949
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1975..2079
                     /label="AmpR promoter"
     CDS             2080..2937
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3111..3699
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3987..4008
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4023..4053
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        4122..4140
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4168..4394
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4395..4575
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4622..4747
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5240..5473
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5658..5702
                     /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        5877..6377
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             6389..7411
                     /label="HygR"
                     /note="hygromycin B phosphotransferase"
     misc_feature    7465..7582
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        7699..8002
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8003..8206
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             8343..9059
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     CDS             9078..10034
                     /label="TurboID"
                     /note="mutant of the E. coli biotin ligase BioID engineered
                     for improved catalytic efficiency (Branon et al., 2018)"
ORIGIN
        1 gtaaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
       61 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
      121 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
      181 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
      241 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
      301 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
      361 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
      421 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
      481 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
      541 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
      601 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
      661 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
      721 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      781 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      841 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
      901 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt
      961 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     1021 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1081 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     1141 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1201 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     1261 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     1321 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1381 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1441 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     1501 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1561 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1621 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1681 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1741 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1801 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1861 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1921 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1981 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2041 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2101 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2161 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2221 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2281 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2341 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2401 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2461 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2521 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2581 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2641 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2701 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2761 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2821 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2881 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2941 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3001 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3061 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3121 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3181 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3241 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3361 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3421 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3481 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3541 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3601 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3661 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3721 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3781 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3841 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3901 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3961 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4021 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4081 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     4141 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     4201 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     4261 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     4321 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     4381 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     4441 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     4501 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     4561 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     4621 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     4681 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     4741 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     4801 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     4861 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     4921 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     4981 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     5041 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     5101 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     5161 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     5221 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     5281 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     5341 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     5401 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     5461 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     5521 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     5581 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     5641 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     5701 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     5761 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     5821 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccttgc
     5881 gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg
     5941 ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca
     6001 cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa
     6061 gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc
     6121 tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat
     6181 gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg
     6241 cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt
     6301 ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca
     6361 ccgacctctc tccccagggg gtaccaccat gaaaaagcct gaactcaccg ctacctctgt
     6421 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctccgaggg
     6481 cgaagaatct cgggctttca gcttcgatgt gggagggcgt ggatatgtcc tgcgggtgaa
     6541 tagctgcgcc gatggtttct acaaagatcg ctatgtttat cggcactttg catccgccgc
     6601 tctccctatt cccgaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     6661 ctcccgccgt gcacagggtg tcaccttgca agacctgcct gaaaccgaac tgcccgctgt
     6721 tctccagccc gtcgccgagg ccatggatgc catcgctgcc gccgatctta gccagaccag
     6781 cgggttcggc ccattcggac ctcaaggaat cggtcaatac actacatggc gcgatttcat
     6841 ctgcgctatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     6901 tgcctccgtc gcccaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     6961 ccggcacctc gtgcacgccg atttcggctc caacaatgtc ctgaccgaca atggccgcat
     7021 aacagccgtc attgactgga gcgaggccat gttcggggat tcccaatacg aggtcgccaa
     7081 catcttcttc tggaggccct ggttggcttg tatggagcag cagacccgct acttcgagcg
     7141 gaggcatccc gagcttgcag gatctcctcg gctccgggct tatatgctcc gcattggtct
     7201 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggctcaggg
     7261 tcgctgcgac gcaatcgtcc ggtccggagc cgggactgtc gggcgtacac aaatcgcccg
     7321 cagaagcgct gccgtctgga ccgatggctg tgtggaagtg ctcgccgata gtggaaacag
     7381 acgccccagc actcgtccta gggcaaagga tctgcagtaa tgagaattct agatcttgag
     7441 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     7501 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     7561 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
     7621 ccggctcgag ggggcccggg gttattaata gtaatcaatt acggggtcat tagttcatag
     7681 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7741 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7801 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7861 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     7921 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     7981 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     8041 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     8101 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     8161 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactgt
     8221 cgggatccac catggagaca gacacactcc tgctatgggt actgctgctc tgggttccag
     8281 gttccactgg tgacggcaag cccatcccca accccctgct gggcctggac agcaccggat
     8341 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     8401 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     8461 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     8521 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     8581 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     8641 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     8701 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     8761 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     8821 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     8881 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     8941 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     9001 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     9061 ggggaggcgg gggtagtaaa gacaatactg tgcctctgaa gctgatcgct ctcctggcta
     9121 atggcgagtt ccatagtggc gaacagctgg gagaaaccct gggcatgtcc agggccgcta
     9181 tcaacaagca cattcagact ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa
     9241 agggctactc tctgcccgag cctatcccgc tgctgaacgc taaacagatt ctgggacagc
     9301 tggacggcgg gagcgtggca gtcctgcctg tggtcgactc caccaatcag tacctgctgg
     9361 atcgaatcgg cgagctgaag agtggggatg cttgcattgc agaatatcag caggcaggga
     9421 gaggaagcag agggaggaaa tggttctctc cttttggagc taacctgtac ctgagtatgt
     9481 tttggcgcct gaagcgggga ccagcagcaa tcggcctggg cccggtcatc ggaattgtca
     9541 tggcagaagc gctgcgaaag ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc
     9601 tgtatctgca ggatagaaag ctggcaggca tcctggtgga gctggccgga ataacaggcg
     9661 atgctgcaca gatcgtcatt ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa
     9721 gcgtggtcaa tcagggctgg atcacactgc aggaagcagg gattaacctg gacaggaata
     9781 ctctggccgc tacgctgatc cgagagctgc gggcagccct ggaactgttc gagcaggaag
     9841 gcctggctcc atatctgcca cggtgggaga agctggataa cttcatcaat agacccgtga
     9901 agctgatcat tggggacaaa gagattttcg ggattagccg ggggattgat aaacagggag
     9961 ccctgctgct ggaacaggac ggagttatca aaccctggat gggcggagaa atcagtctgc
    10021 ggtctgccga aaagggtgga ggcgggtctg gaggcggggg tagtaccggt aaggacgagc
    10081 t
//
