LOCUS       .                       8916 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        320..367
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             386..781
                     /label="BSD"
                     /note="blasticidin S deaminase"
     LTR             871..1104
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1176..1310
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1337..1472
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1493..1511)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1521..1537)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      1679..2134
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2160..2264
                     /label="AmpR promoter"
     CDS             2265..3122
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3296..3884
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4172..4193
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4208..4238
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4246..4262
                     /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)."
     primer_bind     4270..4286
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4307..4325
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4353..4579
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4580..4760
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4807..4932
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5425..5658
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5843..5887
                     /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             6036..6077
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     enhancer        6232..6534
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6535..6738
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        6783..6801
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6889..7013
                     /label="attR1"
                     /note="recombination site for the Gateway® LR reaction"
     promoter        7038..7068
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             7122..7778
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             8123..8425
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(8469..8593)
                     /label="attR2"
                     /note="recombination site for the Gateway® LR reaction"
     CDS             8595..8621
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             8625..8651
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             8655..8681
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     promoter        8723..9222
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
       61 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
      121 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
      181 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
      241 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
      301 ctttcgactc tagacacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
      361 atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct
      421 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
      481 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
      541 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
      601 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
      661 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
      721 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
      781 ctaagcacaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
      841 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
      901 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
      961 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     1021 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     1081 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     1141 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     1201 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1261 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     1321 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     1441 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     1501 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1681 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1741 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1801 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1861 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1921 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1981 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2041 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2101 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3901 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3961 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4021 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4081 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4141 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4201 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4261 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     4321 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     4381 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     4441 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     4501 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     4561 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     4621 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4681 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4741 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     4801 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     4861 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     4921 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     4981 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     5041 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     5101 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     5161 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     5221 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     5281 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     5341 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     5401 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     5461 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     5521 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     5581 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     5641 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     5701 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     5761 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     5821 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     5881 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     5941 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     6001 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     6061 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     6121 attagtgaac ggatctcgac ggtatcgatg tcgacgttaa cgctagcgac tgggtctgcg
     6181 atgtcgacgt taacgctagc gactgggtct gcgataagct tgggagttcc gcgttacata
     6241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6481 atgggacttt cctacttggc agtacatcta gtattagtca tcgctattac catggtgatg
     6541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     6661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     6721 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     6781 attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttggta
     6841 ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatcaac aagtttgtac
     6901 aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     6961 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggcggccg
     7021 cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga
     7081 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     7141 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     7201 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     7261 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     7321 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     7381 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     7441 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     7501 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     7561 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     7621 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     7681 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     7741 aattacaaca gtactgcgat gagtggcagg gcggggcgta aagatctgga tccggcttac
     7801 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
     7861 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt
     7921 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt
     7981 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg
     8041 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct
     8101 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg
     8161 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
     8221 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     8281 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     8341 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     8401 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc
     8461 aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     8521 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
     8581 tgtacaaagt ggtttaccca tacgacgtac ctgattatgc aacttatcct tatgatgtac
     8641 ctgattatgc tagctatcct tatgatgtcc cagactacgc atagtgaacg cgtaccggtt
     8701 agtaatgacg atggtaccta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     8761 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     8821 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     8881 ccttctactc ctcccctagt caggaagttc cccccc
//
