LOCUS       .                      10055 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        7..507
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             529..1125
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             1256..1489
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1561..1695
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1722..1857
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1878..1896)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2064..2519
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2545..2649
                     /label="AmpR promoter"
     CDS             2650..3507
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3681..4269
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4557..4578
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4593..4623
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        4692..4710
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4738..4964
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    5192..5317
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5810..6043
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             6228..6272
                     /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             6421..6462
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     promoter        6570..6810
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        6837..7140
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7141..7344
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             7382..8047
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             8117..8170
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             8195..9844
                     /label="luciferase"
                     /note="firefly luciferase"
     misc_feature    9886..10002
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        10000..10507
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat gaccgagtac
      541 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
      601 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
      661 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
      721 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
      781 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
      841 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
      901 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
      961 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     1021 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     1081 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     1141 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc tttaagacca
     1201 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     1261 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     1321 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     1381 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     1441 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     1501 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     1561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1681 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     1741 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1801 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1861 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     1921 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1981 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2041 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2101 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2161 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2221 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2281 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2341 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2401 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2461 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     2521 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2581 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2641 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2701 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2761 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2821 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2881 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2941 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3001 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3061 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3121 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3181 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3241 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3301 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3361 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3421 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3481 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3541 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3601 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3661 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3721 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3781 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3841 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3901 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3961 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4021 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4081 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4141 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4201 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4261 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4321 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4381 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4441 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4501 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4561 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4621 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     4681 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     4741 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     4801 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     4861 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     4921 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     4981 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5041 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5101 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     5161 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     5221 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     5281 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     5341 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     5401 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     5461 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     5521 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     5581 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     5641 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     5701 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     5761 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     5821 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     5881 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     5941 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     6001 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     6061 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     6121 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     6181 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     6241 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     6301 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     6361 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     6421 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     6481 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag
     6541 actagcctcg agcggccgcc cccttcaccg agggcctatt tcccatgatt ccttcatatt
     6601 tgcatatacg atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa
     6661 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
     6721 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
     6781 tcttggcttt atatatcttg tggaaaggac gaaacaccgg ttttttcgga attccgcgtt
     6841 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6901 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6961 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7021 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7081 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     7141 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     7201 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     7261 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     7321 tgggaggtct atataagcag agctaatcgt cgacgccacc atggagagcg acgagagcgg
     7381 cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga
     7441 gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag
     7501 caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt
     7561 ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa
     7621 cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag
     7681 cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg
     7741 cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga
     7801 gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct
     7861 gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat
     7921 ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct
     7981 gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc
     8041 cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg
     8101 ctccaccgga tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     8161 tcccggccct gattcgtcag aatcggtcgc caccatggaa gacgccaaaa acataaagaa
     8221 aggcccggcg ccattctatc ctcttgagga tggaaccgct ggagagcaac tgcataaggc
     8281 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
     8341 gaacatcacg tacgcggaat acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
     8401 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
     8461 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
     8521 taatgaacgt gaattgctca acagtatgaa catttcgcag cctaccgtag tgtttgtttc
     8581 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaaa ttaccaataa tccagaaaat
     8641 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     8701 atctcatcta cctcccggtt ttaatgaata cgattttgta ccagagtcct ttgatcgtga
     8761 caaaacaatt gcactgataa tgaactcctc tggatctact gggttaccta agggtgtggc
     8821 ccttccgcat agaactgcct gcgtcagatt ctcgcatgcc agagatccta tttttggcaa
     8881 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     8941 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     9001 agaagagctg tttttacgat cccttcagga ttacaaaatt caaagtgcgt tgctagtacc
     9061 aaccctattt tcattcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     9121 acacgaaatt gcttctgggg gcgcacctct ttcgaaagaa gtcggggaag cggttgcaaa
     9181 acgcttccat cttccaggga tacgacaagg atatgggctc actgagacta catcagctat
     9241 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     9301 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc agagaggcga
     9361 attatgtgtc agaggaccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     9421 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     9481 cgaacacttc ttcatagttg accgcttgaa gtctttaatt aaatacaaag gataccaggt
     9541 ggcccccgct gaattggagt cgatattgtt acaacacccc aacatcttcg acgcgggcgt
     9601 ggcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     9661 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     9721 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     9781 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagtccaa
     9841 attgtagttc tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     9901 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     9961 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
    10021 cagcagagat ccactttggc cgcggctcga ggggg
//
