LOCUS       V030218                 9878 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030218
VERSION     V030218
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4710
                     /label="HSP90B1(NM_003299)"
                     /note="HSP90B1(NM_003299)"
                     /gene="HSP90B1"
     CDS             4717..4743
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4747..4773
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4777..4803
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4848..5059
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5072..5340
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5367..5966
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5973..6561
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6635..6868
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6940..7061
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7080..7215
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7253..7269)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7277..7293
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /label="lac operator"
     promoter        complement(7301..7331)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7346..7367
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7655..8243)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8414..9274)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9275..9379)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9853..9869
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
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 gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgagggcc ctgtgggtgc tgggcctctg ctgcgtcctg
     2341 ctgaccttcg ggtcggtcag agctgacgat gaagttgatg tggatggtac agtagaagag
     2401 gatctgggta aaagtagaga aggatcaagg acggatgatg aagtagtaca gagagaggaa
     2461 gaagctattc agttggatgg attaaatgca tcacaaataa gagaacttag agagaagtcg
     2521 gaaaagtttg ccttccaagc cgaagttaac agaatgatga aacttatcat caattcattg
     2581 tataaaaata aagagatttt cctgagagaa ctgatttcaa atgcttctga tgctttagat
     2641 aagataaggc taatatcact gactgatgaa aatgctcttt ctggaaatga ggaactaaca
     2701 gtcaaaatta agtgtgataa ggagaagaac ctgctgcatg tcacagacac cggtgtagga
     2761 atgaccagag aagagttggt taaaaacctt ggtaccatag ccaaatctgg gacaagcgag
     2821 tttttaaaca aaatgactga agcacaggaa gatggccagt caacttctga attgattggc
     2881 cagtttggtg tcggtttcta ttccgccttc cttgtagcag ataaggttat tgtcacttca
     2941 aaacacaaca acgataccca gcacatctgg gagtctgact ccaatgaatt ttctgtaatt
     3001 gctgacccaa gaggaaacac tctaggacgg ggaacgacaa ttacccttgt cttaaaagaa
     3061 gaagcatctg attaccttga attggataca attaaaaatc tcgtcaaaaa atattcacag
     3121 ttcataaact ttcctattta tgtatggagc agcaagactg aaactgttga ggagcccatg
     3181 gaggaagaag aagcagccaa agaagagaaa gaagaatctg atgatgaagc tgcagtagag
     3241 gaagaagaag aagaaaagaa accaaagact aaaaaagttg aaaaaactgt ctgggactgg
     3301 gaacttatga atgatatcaa accaatatgg cagagaccat caaaagaagt agaagaagat
     3361 gaatacaaag ctttctacaa atcattttca aaggaaagtg atgaccccat ggcttatatt
     3421 cactttactg ctgaagggga agttaccttc aaatcaattt tatttgtacc cacatctgct
     3481 ccacgtggtc tgtttgacga atatggatct aaaaagagcg attacattaa gctctatgtg
     3541 cgccgtgtat tcatcacaga cgacttccat gatatgatgc ctaaatacct caattttgtc
     3601 aagggtgtgg tggactcaga tgatctcccc ttgaatgttt cccgcgagac tcttcagcaa
     3661 cataaactgc ttaaggtgat taggaagaag cttgttcgta aaacgctgga catgatcaag
     3721 aagattgctg atgataaata caatgatact ttttggaaag aatttggtac caacatcaag
     3781 cttggtgtga ttgaagacca ctcgaatcga acacgtcttg ctaaacttct taggttccag
     3841 tcttctcatc atccaactga cattactagc ctagaccagt atgtggaaag aatgaaggaa
     3901 aaacaagaca aaatctactt catggctggg tccagcagaa aagaggctga atcttctcca
     3961 tttgttgagc gacttctgaa aaagggctat gaagttattt acctcacaga acctgtggat
     4021 gaatactgta ttcaggccct tcccgaattt gatgggaaga ggttccagaa tgttgccaag
     4081 gaaggagtga agttcgatga aagtgagaaa actaaggaga gtcgtgaagc agttgagaaa
     4141 gaatttgagc ctctgctgaa ttggatgaaa gataaagccc ttaaggacaa gattgaaaag
     4201 gctgtggtgt ctcagcgcct gacagaatct ccgtgtgctt tggtggccag ccagtacgga
     4261 tggtctggca acatggagag aatcatgaaa gcacaagcgt accaaacggg caaggacatc
     4321 tctacaaatt actatgcgag tcagaagaaa acatttgaaa ttaatcccag acacccgctg
     4381 atcagagaca tgcttcgacg aattaaggaa gatgaagatg ataaaacagt tttggatctt
     4441 gctgtggttt tgtttgaaac agcaacgctt cggtcagggt atcttttacc agacactaaa
     4501 gcatatggag atagaataga aagaatgctt cgcctcagtt tgaacattga ccctgatgca
     4561 aaggtggaag aagagcccga agaagaacct gaagagacag cagaagacac aacagaagac
     4621 acagagcaag acgaagatga agaaatggat gtgggaacag atgaagaaga agaaacagca
     4681 aaggaatcta cagctgaaaa agatgaattg ggatcctacc catacgacgt acctgattat
     4741 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     4801 gcataggcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
     4861 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag
     4921 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     4981 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     5041 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
     5101 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
     5161 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     5221 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     5281 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     5341 caagctgtga ccggcgccta cgctagatga ccgagtacaa gcccacggtg cgcctcgcca
     5401 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5461 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5521 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5581 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5641 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5701 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5761 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5821 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5881 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5941 gcatgacccg caagcccggt gcctgagtcg acaatcaacc tctggattac aaaatttgtg
     6001 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     6061 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     6121 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     6181 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     6241 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     6301 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     6361 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     6421 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     6481 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     6541 ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta
     6601 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     6661 aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     6721 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6781 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6841 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     6901 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     6961 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7021 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     7081 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     7141 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     7201 gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc
     7261 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7321 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7381 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7441 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7501 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7561 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7621 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7681 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7741 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7801 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7861 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7921 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7981 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8041 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     8101 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8161 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8221 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8281 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8341 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8401 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8461 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8521 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8581 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8641 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8701 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8761 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8821 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8881 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8941 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9001 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9061 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9121 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9181 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9241 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9301 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9361 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     9421 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     9481 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     9541 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     9601 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     9661 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     9721 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     9781 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     9841 agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
