LOCUS       .                       8356 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        6..232
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    460..585
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1078..1311
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1496..1540
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1689..1730
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    1807..1923
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             2375..2440
                     /label="3xFLAG"
                     /note="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2499..2710
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             2723..2991
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3047..3712
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3782..3835
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3836..4432
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4436..5023
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5097..5330
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5402..5536
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5563..5698
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5731..5747)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5755..5771)
                     /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(5779..5809)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5824..5845)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6133..6721)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6895..7752)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(7753..7857)
                     /label="AmpR promoter"
     primer_bind     8331..8347
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     1921 tttatcgata ctagttgaaa gaccccacct gtaggtttgg caagttagct taagtaacgc
     1981 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     2041 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     2101 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagcg
     2161 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     2221 ctaaccaatc agtttgcttc ttgcttctgt ttgtgtgctt ctgctccctg agctcaataa
     2281 aagagcccac aacccctcac ttggtgggcc agtcctctga tagactgtgt cccctggata
     2341 cccgtattct agagctagcg aattcgccac catggactac aaagaccatg acggtgatta
     2401 taaagatcat gacatcgatt acaaggatga cgatgacaag ctcgagtgca tggatccgcg
     2461 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     2521 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg
     2581 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     2641 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     2701 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac
     2761 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc
     2821 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc
     2881 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct
     2941 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg
     3001 accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga
     3061 gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg
     3121 agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct
     3181 gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac
     3241 ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa
     3301 cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta
     3361 cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag
     3421 cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat
     3481 gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta
     3541 ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct
     3601 gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga
     3661 gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg
     3721 cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg
     3781 cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac
     3841 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3901 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3961 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4021 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4081 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4141 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4201 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4261 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4321 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4381 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca
     4441 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4501 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4561 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4621 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4681 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4741 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4801 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     4861 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4921 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct
     4981 tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac ctttaagacc
     5041 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     5101 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg
     5161 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     5221 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     5281 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     5341 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     5401 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5521 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     5581 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5641 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5701 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     5761 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5821 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5881 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5941 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6001 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6061 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6121 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6181 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6241 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6301 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6361 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6421 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6481 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6541 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6601 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6661 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6721 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6781 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6841 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6901 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6961 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7021 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7081 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7141 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7201 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7261 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7321 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7381 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7441 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7501 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7561 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7621 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7681 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7741 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7801 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7861 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7921 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     8041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     8101 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     8161 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     8221 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     8281 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     8341 ggccagtgcc aagctg
//
