LOCUS       .                       9818 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"
     protein_bind    1998..2268
                     /label="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2301..2339
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2493..3167
                     /label="mRFP1"
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     CDS             3213..3929
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     promoter        3951..4461
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4471..5067
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     CDS             5068..5121
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5122..5865
                     /label="rtTA-Advanced"
                     /note="improved tetracycline-controlled transactivator"
     misc_feature    5913..6501
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6575..6808
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6880..7014
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7020..7155
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(7193..7209)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7217..7233)
                     /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(7241..7271)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7286..7307)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7595..8183)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8357..9214)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(9215..9319)
                     /label="AmpR promoter"
     primer_bind     9793..9809
                     /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 gtggtttatt acagggacag cagagatcca ctttggccgc gaatcgatat
     1981 gtcctcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     2041 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2101 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
     2161 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
     2221 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     2281 gagctcggta cccgggtcga ggtaggcgtg tacggtggga ggcctatata agcagagctc
     2341 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
     2401 gacaccggga ccgatccagc ctccgcggcc ccgaattcat gaagttatgc atattactgg
     2461 ccgtcgtggc ctttgttggc ctctcgctcg ggatggcctc ctccgaggac gtcatcaagg
     2521 agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa cggccacgag ttcgagatcg
     2581 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2641 agggcggccc cctgcccttc gcctgggaca tcctgtcccc tcagttccag tacggctcca
     2701 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     2761 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     2821 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     2881 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgga
     2941 tgtaccccga ggacggcgcc ctgaagggcg agatcaagat gaggctgaag ctgaaggacg
     3001 gcggccacta cgacgccgag gtcaagacca cctacatggc caagaagccc gtgcagctgc
     3061 ccggcgccta caagaccgac atcaagctgg acatcacctc ccacaacgag gactacacca
     3121 tcgtggaaca gtacgagcgc gccgagggcc gccactccac cggcgccatc gatgaattca
     3181 ctagtgttaa cggatccctc gagcggccgc ccatggtgag caagggcgag gagctgttca
     3241 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3301 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3361 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     3421 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3481 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3541 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3601 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3661 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3721 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3781 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     3841 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3901 tcactctcgg catggacgag ctgtacaaga aagatgagct ctaaggatcc ggggttgggg
     3961 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     4021 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     4081 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     4141 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     4201 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     4261 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     4321 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     4381 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     4441 atcaccgacc tctctcccca gcaattcacc atgaccgagt acaagcccac ggtgcgcctc
     4501 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4561 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4621 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4681 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4741 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4801 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4861 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     4921 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4981 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5041 tggtgcatga cccgcaagcc cggtgccgaa ggtagaggtt ctctcctcac ttgtggtgat
     5101 gttgaagaaa accctggtcc aatgtctaga ctggacaaga gcaaagtcat aaacggagct
     5161 ctggaattac tcaatggtgt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag
     5221 ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat
     5281 gccctgccaa tcgagatgct ggacaggcat catacccact tctgccccct ggaaggcgag
     5341 tcatggcaag actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat
     5401 cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg
     5461 gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct
     5521 ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta
     5581 gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct gagacaagca
     5641 attgagctgt tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc
     5701 atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac
     5761 gattttgact tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg
     5821 cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccgggtaact aagtaaggat
     5881 cgatccaaga tatcgtattc ttaactgtcg acaatcaacc tctggattac aaaatttgtg
     5941 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     6001 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     6061 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     6121 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     6181 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     6241 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     6301 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     6361 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     6421 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     6481 ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta
     6541 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     6601 aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     6661 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6721 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6781 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     6841 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     6901 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6961 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     7021 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     7081 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     7141 gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc
     7201 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7261 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7321 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7381 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7441 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7501 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     8041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8941 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     9361 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     9421 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     9481 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     9541 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     9601 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     9661 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     9721 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     9781 agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
