LOCUS       .                      10233 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        237..616
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..819
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    1061..1186
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1683..1916
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2101..2145
                     /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             2294..2335
                     /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    2443..2560
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2617..3795
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
     enhancer        3907..4192
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV
                     enhancer"
     promoter        4194..4471
                     /label="chicken β-actin promoter"
     CDS             4726..6375
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             6385..6441
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             6448..7044
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    7122..7710
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8239..8419
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8481..9069)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9243..10100)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(10101..10205)
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gcatgcggct ccggtgcccg tcagtgggca
     2641 gagcgcacat cgcccacagt ccccgagaag ttgtggggag gggtcggcaa ttgaaccggt
     2701 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     2761 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     2821 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
     2881 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
     2941 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
     3001 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
     3061 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
     3121 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
     3181 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
     3241 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     3301 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
     3361 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     3421 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     3481 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     3541 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     3601 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     3661 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     3721 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     3781 tttcaggtgt cgtgaggaat cctctaggtc gacctggatc ccgatacgac ctcgagctcc
     3841 cggaattcgt cgacgcggcc gcatctctgc gacgcgtcga atcgtcgcta gcgcctctag
     3901 ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3961 cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4021 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4081 tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg
     4141 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     4201 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     4261 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc
     4321 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     4381 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     4441 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg
     4501 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     4561 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
     4621 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
     4681 ctgaaatcac tttttttcag gttggaccgg tctaggcgcg ccaccatgga agacgccaaa
     4741 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
     4801 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
     4861 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
     4921 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     4981 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     5041 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     5101 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     5161 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     5221 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     5281 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
     5341 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     5401 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     5461 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     5521 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     5581 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     5641 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     5701 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     5761 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     5821 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     5881 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     5941 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     6001 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     6061 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     6121 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     6181 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     6241 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     6301 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     6361 ggaaagatcg ccgtgggaag cggagctact aacttcagcc tgctgaagca ggctggagac
     6421 gtggaggaga accctggacc tgtcgacatg accgagtaca agcccacggt gcgcctcgcc
     6481 acccgcgacg acgtcccccg ggcagtacgc accctcgccg ccgcgttcgc cgactacccc
     6541 gccacgcgcc acaccgtcga tccagaccgc cacatcgagc gggtcaccga gctgcaagaa
     6601 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6661 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6721 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6781 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     6841 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6901 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6961 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     7021 tgcatgaccc gcaagcccgg tgcctgagct ttgtttaaac ggcgcgacat gtttaagggt
     7081 tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct ctggattaca
     7141 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     7201 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     7261 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     7321 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     7381 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     7441 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     7501 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     7561 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     7621 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     7681 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgagaccta
     7741 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     7801 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     7861 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     7921 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     7981 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg
     8041 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat
     8101 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     8161 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
     8221 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     8281 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     8341 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     8401 gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     8941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     9001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     9061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     9121 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     9181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     9241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     9301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     9361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     9421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     9481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     9541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     9601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     9661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    10021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    10081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    10141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc
    10201 ccgcgcacat ttccccgaaa agtgccacct gac
//
