LOCUS       V032928                 8803 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032928
VERSION     V032928
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..3339
                     /label="Arf6(NM_007481)"
                     /note="Arf6(NM_007481)"
                     /gene="Arf6"
     CDS             3340..3366
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3377..3963
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             3964..4758
                     /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    4772..5360
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5567..6200
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6328..6344)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6352..6368
                     /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(6376..6406)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6421..6442
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6730..7318)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7489..8349)
                     /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(8350..8454)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8502..8636
                     /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 caccatgggg
     2821 aaggtgctat ccaagatctt cgggaacaag gaaatgcgga tcctcatgct gggcctggac
     2881 gcagccggca agacaacgat cctgtacaag ttgaagctgg gccaatcggt gaccaccatc
     2941 cccacggtgg gcttcaacgt ggagacggtg acttacaaaa acgtcaagtt caacgtgtgg
     3001 gatgtgggcg gccaggacaa gatccggccg ctctggcggc attactacac cgggacccag
     3061 ggtctgatct tcgtggtaga ctgcgccgac cgcgaccgca tcgacgaggc ccgccaggag
     3121 ctgcaccgca ttatcaatga ccgggagatg agggacgcca tcatcctcat cttcgccaac
     3181 aagcaggacc tgcccgatgc catgaaaccc catgagatcc aggagaaact gggcctgacc
     3241 cggattcggg acaggaactg gtatgtgcag ccctcctgtg ccacctccgg ggacggactc
     3301 tatgaggggc tcacatggtt aacctctaac tacaaatcct acccatacga tgttccagat
     3361 tacgcttagg gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     3421 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     3481 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     3541 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     3601 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     3661 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     3721 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     3781 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     3841 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     3901 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca
     3961 accatggctg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4021 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4081 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4141 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4201 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4261 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4321 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4381 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4441 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4501 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4561 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4621 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4681 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4741 cttgacgagt tcttctgaac gcgtctggaa caatcaacct ctggattaca aaatttgtga
     4801 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4861 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4921 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4981 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5041 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5101 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5161 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5221 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5281 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5341 cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga
     5401 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     5461 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     5521 aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt
     5581 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     5641 cctgattagc agaactacac accagggcca ggggtcagat atccactgac ctttggatgg
     5701 tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac
     5761 accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta
     5821 gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag
     5881 tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca
     5941 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca
     6001 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6061 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6121 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6181 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6241 tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt taccgtcgac
     6301 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7141 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8461 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt
     8521 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     8581 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
     8641 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
     8701 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     8761 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
