LOCUS       V022497                 9903 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022497
VERSION     V022497
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2815..4374
                     /label="USP3(NM_006537)"
                     /note="USP3(NM_006537)"
                     /gene="USP3"
     CDS             4381..4446
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4477..5063
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5064..5858
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5872..6460
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6667..7300
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7428..7444)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7452..7468
                     /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(7476..7506)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7521..7542
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7830..8418)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8589..9449)
                     /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(9450..9554)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9602..9736
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggag
     2821 tgtccacacc tgagctccag cgtctgcatt gctccggact cagccaagtt ccccaacggc
     2881 tccccgtcgt cctggtgctg cagcgtgtgc cggtccaaca aaagcccttg ggtctgtttg
     2941 acttgttcaa gtgtccactg tggaaggtat gtgaatggcc atgcaaaaaa acattatgaa
     3001 gatgcacaag tacctttaac caaccataag aaatcagaaa agcaagataa agttcagcac
     3061 acagtatgta tggattgcag tagctacagt acatactgtt atcgctgtga tgattttgtg
     3121 gttaatgaca ccaagctggg actggtacag aaagtcagag aacacttaca gaacttggaa
     3181 aactcagctt tcacagctga caggcataag aaaagaaaac ttttggaaaa ctcaacacta
     3241 aacagcaagt tattaaaagt aaatggaagc accactgcca tttgtgccac aggccttcgg
     3301 aatttgggga acacatgttt catgaatgcc atccttcagt cactcagtaa cattgagcag
     3361 ttttgctgtt atttcaaaga actgcccgcc gtggagttaa ggaatgggaa aacagcagga
     3421 aggcggacat accacaccag gagccaaggg gataacaatg tgtctttggt agaagagttt
     3481 agaaagacac tctgtgcttt atggcaaggc agccagactg catttagccc agagtcctta
     3541 ttttatgttg tttggaagat tatgccaaac tttaggggct atcaacagca ggacgcccat
     3601 gaattcatgc gctacctttt ggaccaccta cacttggaac ttcagggcgg tttcaacggt
     3661 gtttcccgct cagcaattct gcaggagaat tctactctgt ctgcaagtaa caagtgttgc
     3721 ataaatggag catctactgt tgtcacggct atattcggag gcattctcca aaatgaggtt
     3781 aactgcctca tatgtgggac agaatctaga aagtttgatc cattcctaga cctttcatta
     3841 gatattccaa gtcagttcag aagtaagcgc tctaagaatc aagaaaatgg accagtttgt
     3901 tcgttacgag attgtcttcg cagttttacc gacttagaag aacttgatga gacagagtta
     3961 tatatgtgcc ataaatgcaa aaagaaacaa aagtccacaa aaaagttttg gattcaaaaa
     4021 ctacccaagg tgctatgctt acatttgaaa agatttcatt ggacagcata tttaagaaat
     4081 aaagttgata catacgtaga atttccactg agaggcctag acatgaaatg ctacttacta
     4141 gagcctgaga acagtggccc ggagagctgc ctgtatgacc tcgccgctgt ggtggtgcac
     4201 catggttccg gggttggttc tggacattac acagcatacg caactcacga aggccgctgg
     4261 ttccacttca atgacagtac tgtaacactg actgacgagg agactgtggt gaaggcgaag
     4321 gcctacatcc ttttctacgt ggaacaccag gccaaagctg gatcggataa acttctcgag
     4381 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4441 gacaagtaga ctagttctag agcggccgcg gatccgcccc tctccctccc ccccccctaa
     4501 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     4561 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     4621 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     4681 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     4741 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4801 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4861 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4921 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     4981 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     5041 acgatgataa gcttgccaca accatggctg aacaagatgg attgcacgca ggttctccgg
     5101 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5161 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5221 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5281 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5341 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5401 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5461 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5521 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5581 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5641 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5701 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5761 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5821 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaac gcgtctggaa caatcaacct
     5881 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5941 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     6001 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     6061 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6121 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6181 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6241 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     6301 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6361 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     6421 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc
     6481 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     6541 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     6601 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga
     6661 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     6721 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat
     6781 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag
     6841 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg
     6901 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg
     6961 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac
     7021 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc
     7081 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag
     7141 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     7201 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7261 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     7321 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat
     7381 tgctagcgtt taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     7441 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7501 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7621 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7681 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7741 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7801 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7861 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7921 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7981 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     8041 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8101 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8161 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8221 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     8281 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8341 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8821 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8881 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8941 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9001 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9061 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9121 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9181 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9241 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     9301 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     9361 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     9421 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     9481 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9541 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     9601 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     9661 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     9721 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac
     9781 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc
     9841 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     9901 agt
//
