LOCUS       V043520                 9803 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043520
VERSION     V043520
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             2290..4203
                     /label="Uba2(NM_016682)"
                     /note="Uba2(NM_016682)"
                     /gene="Uba2"
     CDS             4210..4236
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4240..4266
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4270..4296
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4346..4557
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4570..4838
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4866..5891
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5898..6486
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6560..6793
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6865..6986
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7005..7140
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7178..7194)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7202..7218
                     /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(7226..7256)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7271..7292
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7580..8168)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8339..9199)
                     /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(9200..9304)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9778..9794
                     /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 agagccacca tggcactgtc gcgggggttg ccccgggagc tggccgaggc ggtgtccggc
     2341 ggccgggtgc tggtggttgg cgcgggtggg atcggctgtg agctgctcaa gaacctggtg
     2401 ctcactggct tctcccacat cgacctgatt gatctggaca ctatcgatgt cagcaacctc
     2461 aacaggcagt tcttgtttca aaagaaacat gtgggaagat caaaggctca ggttgccaaa
     2521 gaaagtgtgc tccagttcca cccccaagcc aacattgagg ctcaccatga cagcatcatg
     2581 aacccagact ataatgtgga gttttttcga cagtttatat tggtcatgaa tgcattagat
     2641 aacagagctg cacgaaacca cgtgaatagg atgtgtctgg ccgctgatgt gcctctcatt
     2701 gagagcggga ctgctgggta tctcggacag gtgaccacta tcaagaaggg tgtgactgag
     2761 tgttacgaat gtcaccctaa gcctacccag aggacgttcc ctggctgtac gattcggaac
     2821 acaccttcag aacctatcca ttgcatcgta tgggccaagt atttgttcaa ccagctgttt
     2881 ggagaggaag atgctgatca agaagtgtct cctgacagag ctgaccctga ggctgcttgg
     2941 gaaccaacag aggctgaagc tagagctcga gcatctaatg aagacggaga cattaagcgt
     3001 atttccacta aggaatgggc taagtcaact ggatatgatc cagttaaact ttttaccaag
     3061 ctttttaaag atgatattag atatctgctg acaatggaca aactctggcg caagaggaaa
     3121 cctccagttc cactggactg ggctgaagtg cagagtcaag gagaagccaa tgccgatcaa
     3181 caaaatgagc cccagttggg tttgaaagac cagcaggttc tggatgtaaa gagctatgcg
     3241 agtctgtttt caaagagcat cgagactctg cgagtccatt tagcggagaa gggggatgga
     3301 gctgagctca tctgggataa ggacgaccca cctgcaatgg attttgttac atctgctgca
     3361 aacctcagga tgcacatttt cagcatgaat atgaagagca ggtttgatat aaaatcaatg
     3421 gcagggaata tcattcctgc tatagctact actaatgcag tgatcgccgg cttgatagtg
     3481 ttggaaggat taaagatttt atctggaaaa atagaccagt gtagaactat ttttttgaat
     3541 aagcagccaa acccaagaaa gaagctcttg gtgccctgtg cactggatcc tcccaacacc
     3601 aactgttacg tttgtgccag caagccagag gtgactgtgc ggctgaatgt ccacaaagtg
     3661 actgtcctca ccttacagga taagatagtg aaagaaaagt ttgctatggt ggcaccagat
     3721 gtccaaattg aagatgggaa aggcacaatc ctgatatctt cagaagaggg agagacagaa
     3781 gccaataacc ccaagaagtt gtctgatttt ggaattagaa acggcagccg gcttcaagca
     3841 gacgacttcc tccaggacta tactttgctg atcaacattc ttcatagtga agatcttgga
     3901 aaggatgttg aatttgaagt agttggtgac agccctgaaa aagtggggcc aaaacaagct
     3961 gaagatgctg ccaaaagcat tgccaacggc agcgatgacg gtgcccagcc ttccacctcc
     4021 acagcacaag agcaagatga tgtgctcata gttgattcag acgaagaagg cccttcaaat
     4081 agcaccgact gcagcgggga cgacaaggcc cgcaagcgga agctggaaga gaatgaggct
     4141 gccagtacca agaagtgccg cttggagcag atggaagatc cagatgacgt catagcgtta
     4201 gacctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat
     4261 tatgctagct atccttatga tgtcccagac tacgcatagg gatccgcggc cgcaaggatc
     4321 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4381 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     4441 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4501 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     4561 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     4621 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     4681 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     4741 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     4801 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     4861 tcgacatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4921 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4981 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     5041 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     5101 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     5161 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     5221 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     5281 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     5341 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     5401 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     5461 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     5521 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5581 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5641 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5701 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5761 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     5821 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5881 caaaggaata ggtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5941 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     6001 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6061 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6121 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6181 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6241 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6301 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6361 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6421 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6481 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6541 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt
     6601 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6661 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6721 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6781 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6841 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6901 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6961 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     7021 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7081 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7141 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     7201 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7261 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7321 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7381 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7441 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7501 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7561 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7621 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7981 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     8041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9301 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9361 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     9421 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     9481 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9541 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9601 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     9661 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     9721 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     9781 aaacgacggc cagtgccaag ctg
//
