LOCUS       .                       9248 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      46..474
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      647..782
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        844..891
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             910..1281
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    1414..1547
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1584..1600)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1608..1624)
                     /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(1632..1662)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1677..1698)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1986..2574)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2748..3605)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(3606..3710)
                     /label="AmpR promoter"
     enhancer        3976..4355
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4460..4558
                     /label="CMVd1 promoter"
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields slightly reduced expression
                     relative to the full-length promoter"
     misc_feature    4800..4925
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5418..5651
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5836..5880
                     /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             6029..6070
                     /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    6178..6295
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        6388..6599
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     CDS             6624..7340
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     misc_feature    7830..8418
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(8421..8437)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        8595..8797
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             8812..8992
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    9024..9248
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
       61 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      121 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      181 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      241 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      301 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      361 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      421 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
      481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      661 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
      721 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
      781 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
      841 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
      901 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
      961 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     1021 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     1081 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     1141 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     1201 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     1261 ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
     1321 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1381 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     1441 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1501 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     1561 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1621 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1681 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1741 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1801 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1861 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1921 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1981 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2041 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2101 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2161 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2221 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2281 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2341 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2401 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2461 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2521 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2581 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2641 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2701 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2761 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2821 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2881 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2941 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3001 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3061 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3121 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3181 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3241 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3301 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3361 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3421 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3481 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3541 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3601 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3661 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3721 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     3781 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     3841 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     3901 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     3961 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     4021 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4081 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4141 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4201 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4261 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4321 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4381 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4441 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4501 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt
     4561 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     4621 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     4681 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     4741 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     4801 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     4861 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     4921 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     4981 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     5041 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     5101 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     5161 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     5221 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     5281 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     5341 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     5401 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     5461 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     5521 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     5581 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     5641 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     5701 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     5761 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     5821 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     5881 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     5941 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     6001 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     6061 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     6121 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
     6181 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     6241 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     6301 ttattacagg gacagcagag atccagtttg gttaattagc tagctaggtc ttgaaaggag
     6361 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     6421 agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact
     6481 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     6541 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     6601 accggttcta gagcgctgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     6661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     6721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     6781 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     6841 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     6901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     6961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     7021 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     7081 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     7141 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     7201 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     7261 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     7321 gcatggacga gctgtacaag ggatccggcg caacaaactt ctctctgctg aaacaagccg
     7381 gagatgtcga agagaatcct ggaccgatgg ccaagccttt gtctcaagaa gaatccaccc
     7441 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     7501 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     7561 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     7621 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     7681 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     7741 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     7801 gctaagaatt cgatatcaag cttatcggta atcaacctct ggattacaaa atttgtgaaa
     7861 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     7921 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     7981 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     8041 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     8101 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     8161 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     8221 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     8281 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     8341 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     8401 tttgggccgc ctccccgcat cgataccgtc gacctcgagg tcgactttaa gaccaatgac
     8461 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     8521 aattcactcc caacgaagac aagatttcac cattatcgtt tcagacccac ctcccaaccc
     8581 cgaggggacc cagagtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     8641 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     8701 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     8761 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
     8821 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     8881 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     8941 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
     9001 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     9061 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     9121 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     9181 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     9241 ctctatgg
//
