LOCUS       .                      10166 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    458..583
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /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             1686..1727
                     /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    1804..1920
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2010..2213
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2302..3018
                     /label="ECFP"
                     /note="enhanced CFP"
     CDS             3070..3093
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     promoter        3136..3347
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             3360..3628
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3687..5336
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             5406..5459
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5460..6251
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    6261..6849
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6923..7156
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7228..7362
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7368..7503
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(7589..7619)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7634..7655)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7943..8531)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8705..9562)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(9563..9667)
                     /label="AmpR promoter"
     primer_bind     10141..10157
                     /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 cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
     2761 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg tggaggcgga tcaggaggcg gtggatccgc tcgaattaaa
     3061 tcactcgagg attacaagga cgacgatgac aagtagcggc cgcaaggatc tgcgatcgct
     3121 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     3181 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     3241 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     3301 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag
     3361 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     3421 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     3481 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     3541 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt
     3601 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca
     3661 ccatggagag cgacgagagc ggcctgatgg aagacgccaa aaacataaag aaaggcccgg
     3721 cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga
     3781 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca
     3841 cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
     3901 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg
     3961 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac
     4021 gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg
     4081 ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca
     4141 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc
     4201 tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa
     4261 ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc
     4321 atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca
     4381 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta
     4441 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc
     4501 tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat
     4561 tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa
     4621 ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc
     4681 atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta
     4741 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga
     4801 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg
     4861 tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga
     4921 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact
     4981 tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg
     5041 ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc
     5101 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga
     5161 cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt
     5221 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg
     5281 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgagat
     5341 cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat
     5401 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta
     5461 tggctgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5521 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5581 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5641 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5701 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5761 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5821 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5881 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5941 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     6001 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6061 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6121 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6181 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6241 acgagttctt ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     6301 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6361 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6421 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6481 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6541 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6601 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6661 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6721 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6781 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6841 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6901 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg
     6961 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7021 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7081 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7141 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     7201 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     7261 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7321 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     7381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     7441 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7501 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt
     7561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7681 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7741 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7801 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7861 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8341 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     8401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8701 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8761 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8821 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8881 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8941 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9001 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9061 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9121 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9181 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9241 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9301 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9361 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9421 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9481 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9541 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9601 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9661 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9721 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9781 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9841 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     9901 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     9961 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
    10021 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
    10081 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
    10141 gtaaaacgac ggccagtgcc aagctg
//
