LOCUS       .                       9371 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..1921
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2011..2214
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     sig_peptide     2317..2379
                     /label="Igκ leader"
                     /note="leader sequence from mouse immunoglobulin κ light
                     chain"
     CDS             3463..3486
                     /label="8xHis"
                     /note="8xHis affinity tag"
     promoter        3530..3741
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             3754..4022
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4078..4743
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             4813..4866
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4867..5463
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5467..6054
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6128..6361
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6433..6567
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6573..6708
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(6746..6762)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6770..6786)
                     /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(6794..6824)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6839..6860)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7148..7736)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7910..8767)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(8768..8872)
                     /label="AmpR promoter"
     primer_bind     9346..9362
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg agacagacac actcctgcta
     2341 tgggtactgc tgctctgggt tccaggttcc actggtgacc aggttcagct gcaggaatct
     2401 ggtggtggtc tggttcagcc gggtggttct ctgcgtctgt cttgcgctgc ttctggtaaa
     2461 atgtcttctc gtcgttgcat ggcttggttc cgtcaggctc cgggtaaaga acgtgaacgt
     2521 gttgctaaac tgctgaccac ctctggttct acctacctgg ctgactctgt taaaggtcgt
     2581 ttcaccatct ctcagaacaa cgctaaatct accgtttacc tgcagatgaa ctctctgaaa
     2641 ccggaagaca ccgctatgta ctactgcgct gctgactctt tcgaagaccc gacctgcacc
     2701 ctggttacct cttctggtgc tttccagtac tggggtcagg gtacccaggt taccgtttct
     2761 gaaccgaaat cttgcgacaa aacccacacc tgcccgccgt gcccggctcc ggaactgctg
     2821 ggtggtccgt ctgttttcct gttcccgccg aaaccgaaag acaccctgat gatctctcgt
     2881 accccggaag ttacctgcgt tgttgttgac gtttctcacg aagacccgga agttaaattc
     2941 aactggtacg ttgacggtgt tgaagttcac aacgctaaaa ccaaaccgcg tgaagaacag
     3001 tacaactcta cctaccgtgt tgtttctgtt ctgaccgttc tgcaccagga ctggctgaac
     3061 ggtaaagaat acaaatgcaa agtttctaac aaagctctgc cggctccgat cgaaaaaacc
     3121 atctctaaag ctaaaggtca gccgcgtgaa ccgcaggttt acaccctgcc gccgtctcgt
     3181 gacgaactga ccaaaaacca ggtttctctg acctgcctgg ttaaaggttt ctacccgtct
     3241 gacatcgctg ttgaatggga atctaacggt cagccggaaa acaactacaa aaccaccccg
     3301 ccggttctgg actctgacgg ttctttcttc ctgtactcta aactgaccgt tgacaaatct
     3361 cgttggcagc agggtaacgt tttctcttgc tctgttatgc acgaagctct gcacaaccac
     3421 tacacccaga aatctctgtc tctgtctccg ggtaaaggat cccaccatca ccatcaccat
     3481 caccattagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     3541 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     3601 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     3661 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     3721 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     3781 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     3841 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     3901 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     3961 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4021 tccaagctgt gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg
     4081 cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg
     4141 gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc
     4201 aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac
     4261 cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc
     4321 ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc
     4381 agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc
     4441 cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac
     4501 ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc
     4561 gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac
     4621 cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac
     4681 agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc
     4741 gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc
     4801 accggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     4861 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4921 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4981 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5041 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5101 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5161 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     5221 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5281 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     5341 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5401 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5461 gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5521 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5581 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5641 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5701 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5761 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5821 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     5881 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5941 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6001 cgtctccgcc ttcgccctca gacgagtcgg atctcccttt ggccgcctcc ccgcctggta
     6061 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6121 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     6181 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6241 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6301 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6361 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6421 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6481 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6541 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6721 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6781 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6841 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6901 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6961 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7021 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7081 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7141 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7201 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7261 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7321 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7381 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7441 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7501 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7561 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7621 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7681 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7741 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7801 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7861 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7921 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7981 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8041 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8101 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8161 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8221 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8281 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8341 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8401 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8461 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8521 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8581 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8641 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8701 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8761 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8821 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8881 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8941 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     9001 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     9061 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     9121 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     9181 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     9241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     9301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     9361 gtgccaagct g
//
