LOCUS       V043366                 9103 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043366
VERSION     V043366
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3129
                     /label="OTUB1(NM_017670)"
                     /note="OTUB1(NM_017670)"
                     /gene="OTUB1"
     CDS             3136..3201
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3245..3456
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3469..3737
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3766..3775
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3796..4458
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4528..4581
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4582..5181
                     /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    5182..5770
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5844..6077
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6149..6270
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6310..6445
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6478..6494)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6502..6518
                     /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(6526..6556)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6571..6592
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6880..7468)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7639..8499)
                     /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(8500..8604)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9078..9094
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg cggcggagga acctcagcag
     2341 cagaagcagg agccgctggg cagcgactcc gaaggtgtta actgtctggc ctatgatgaa
     2401 gccatcatgg ctcagcagga ccgaattcag caagagattg ctgtgcagaa ccctctggtg
     2461 tcagagcggc tggagctctc ggtcctatac aaggagtatg ctgaagatga caacatctat
     2521 caacagaaga tcaaggacct ccacaaaaag tactcgtaca tccgcaagac caggcctgac
     2581 ggcaactgtt tctatcgggc tttcggattc tcccacttgg aggcactgct ggatgacagc
     2641 aaggagttgc agcggttcaa ggctgtgtct gccaagagca aggaagacct ggtgtcccag
     2701 ggcttcactg aattcacaat tgaggatttc cacaacacgt tcatggacct gattgagcag
     2761 gtggagaagc agacctctgt cgccgacctg ctggcctcct tcaatgacca gagcacctcc
     2821 gactaccttg tggtctacct gcggctgctc acctcgggct acctgcagcg cgagagcaag
     2881 ttcttcgagc acttcatcga gggtggacgg actgtcaagg agttctgcca gcaggaggtg
     2941 gagcccatgt gcaaggagag cgaccacatc cacatcattg cgctggccca ggccctcagc
     3001 gtgtccatcc aggtggagta catggaccgc ggcgagggcg gcaccaccaa tccgcacatc
     3061 ttccctgagg gctccgagcc caaggtctac cttctctacc ggcctggaca ctacgatatc
     3121 ctctacaaag gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3181 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3241 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3301 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3361 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3421 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     3481 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3541 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     3601 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     3661 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     3721 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc
     3781 gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc
     3841 gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac
     3901 aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg
     3961 ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac
     4021 gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg
     4081 ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg
     4141 gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac
     4201 gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc
     4261 accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc
     4321 aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc
     4381 gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag
     4441 acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc
     4501 gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac
     4561 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5041 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5161 acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact
     5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5581 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5761 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5821 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct
     5881 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5941 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6001 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6061 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6121 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6241 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6361 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6421 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc
     6481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6781 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6901 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6961 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7021 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7081 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7141 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7201 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7261 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7321 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7381 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7441 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7501 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7561 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7621 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7801 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8161 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8461 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8521 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8581 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8641 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8701 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8761 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8821 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8881 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9001 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9061 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
