LOCUS       .                       8468 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(join(8382..8468,1..18))
                     /label="AmpR promoter"
     primer_bind     492..508
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        523..750
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    975..1100
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1593..1826
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2010..2054
                     /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             2203..2244
                     /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    2315..2432
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2522..2725
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2877..2900
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2904..2927
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3138..3854
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     CDS             3861..3884
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3888..3911
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     promoter        3955..4166
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             4179..4447
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4474..5070
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5080..5668
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5742..5975
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6047..6181
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6187..6322
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(6360..6376)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6384..6400)
                     /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(6408..6438)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6453..6474)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6762..7350)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7524..8381)
                     /label="AmpR"
                     /note="β-lactamase"
ORIGIN
        1 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
       61 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
      121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      241 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
      361 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      421 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
      481 gtcacgacgt tgtaaaacga cggccagtgc caagctgacg cgtgtagtct tatgcaatac
      541 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
      601 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
      661 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
      721 tatttaagtg cctagctcga tacaataaac gggtctctct ggttagacca gatctgagcc
      781 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      841 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      901 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ctgaaagcga
      961 aagggaaacc agagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1021 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1081 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1141 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1201 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1261 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1321 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1381 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1441 ccactgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     1501 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     1561 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     1621 gcagcaggaa gcactatggg cgcagcctca atgacgctga cggtacaggc cagacaatta
     1681 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     1741 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     1801 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1861 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gattggaatc
     1921 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1981 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2041 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2101 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2161 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2221 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2281 ccattcgatt agtgaacgga tctcgacggt taacttttaa aagaaaaggg gggattgggg
     2341 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2401 tacaaaaaca aattacaaaa attcaaaatt ttatcgatac tagtattatg cccagtacat
     2461 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2521 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     2581 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     2641 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     2701 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     2761 tccacgctgt tttgacctcc atagaagatt ctagaatgaa agaaaccgct gctgctaaat
     2821 tcgaacgcca gcacatggac agcggagccg gcgcaggagc cggcgctgac gcgcctgact
     2881 acaaagacga tgacgacaag ggagattaca aggatgacga tgacaaaggc gcgtcgatgg
     2941 acgagaagac caccggctgg cggggcggcc acgtggtgga gggcctggcc ggcgagctgg
     3001 agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc tccggcggct
     3061 gcaagctggg cggcgcgcct gaattctgca gtcgacggta ccgcgggccc atcgctcgat
     3121 ctagagctag cctcgagatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3181 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3241 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3301 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3361 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3421 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3481 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3541 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3601 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3661 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3721 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3781 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3841 acgagctgta caagggatcc gattataagg atgatgacga caaaggagac tataaagacg
     3901 acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     3961 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     4021 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     4081 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     4141 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     4201 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     4261 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     4321 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     4381 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     4441 acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc
     4501 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4561 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4621 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4681 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4741 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4801 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4861 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4921 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4981 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5041 acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa
     5101 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5161 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5221 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5281 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5341 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5401 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5461 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     5521 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5581 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5641 cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc
     5701 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     5761 ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     5821 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     5881 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     5941 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6001 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6061 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6121 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     6181 ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6241 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6301 tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg
     6361 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6421 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6481 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6541 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6601 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6661 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6721 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6781 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6841 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6901 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6961 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7021 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7081 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7141 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7201 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7261 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7321 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7381 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7441 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7501 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7561 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7621 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7681 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7741 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7801 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7861 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7921 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7981 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8041 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8101 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8161 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8221 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8281 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8341 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8401 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8461 aaaaataa
//
