LOCUS       .                       8203 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..204
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             351..374
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             384..1100
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     promoter        1145..1356
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             1369..1637
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1664..2260
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    2270..2858
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2932..3165
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3237..3371
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3398..3533
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(3566..3582)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3590..3606)
                     /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(3614..3644)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3659..3680)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3968..4556)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4730..5587)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(5588..5692)
                     /label="AmpR promoter"
     primer_bind     6166..6182
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6197..6423
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6424..6604
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6651..6776
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7269..7502
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7687..7731
                     /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             7880..7921
                     /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    7998..8114
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggttt atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagattc tagagctagc gaattcgcca ccgtttagaa
      301 tcctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
      361 atgacgatga caaggggcgc gccatggtga gcaagggcga ggagctgttc accggggtgg
      421 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      481 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      541 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
      601 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      661 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      721 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      781 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
      841 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
      901 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
      961 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1021 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1081 gcatggacga gctgtacaag taagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     1141 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     1201 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     1261 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     1321 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     1381 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     1441 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     1501 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     1561 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     1621 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     1681 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     1741 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     1801 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     1861 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     1921 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     1981 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     2041 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     2101 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     2161 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2221 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     2281 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2341 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2401 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2461 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2521 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2581 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2641 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2701 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2761 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2821 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     2881 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     2941 taattcactc ccaacgaaga taagatctgc tttttgcttg tactgggtct ctctggttag
     3001 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     3061 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     3121 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     3181 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     3241 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3301 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3361 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3481 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     3541 ttgcagagac caaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     3601 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3661 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4381 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5101 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5701 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5761 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     5821 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5881 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     5941 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6001 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     6061 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6181 gtgccaagct gacgcgtgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     6241 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     6301 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     6361 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     6421 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6481 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6541 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6601 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     6661 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     6721 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     6781 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     6841 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     6901 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     6961 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     7021 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     7081 agcaaaacaa aagtaagacc accgcacagc aagcggccac tgatcttcag acctggagga
     7141 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     7201 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     7261 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     7321 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     7381 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     7441 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     7501 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     7561 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7621 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7681 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7741 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     7801 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     7861 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     7921 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     7981 cgacggtatc ggttaacttt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     8041 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     8101 aaattcaaaa ttttatcgat actagtatta tgcccagtac atgaccttat gggactttcc
     8161 tacttggcag tacatctacg tattagtcat cgctattacc atg
//
