LOCUS       V024870                10097 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024870
VERSION     V024870
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"
     promoter        2337..3519
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3551..3568
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             3569..4756
                     /label="GPR84(NM_020370)"
                     /note="GPR84(NM_020370)"
                     /gene="GPR84"
     CDS             4763..4828
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4847..5433
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5453..6052
                     /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    6066..6654
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6861..7494
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7622..7638)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7646..7662
                     /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(7670..7700)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7715..7736
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8024..8612)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8783..9643)
                     /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(9644..9748)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9796..9930
                     /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt
     3541 cgccaccatg caccatcacc atcaccatat gtggaacagc tctgacgcca acttctcctg
     3601 ctaccatgag tctgtgctgg gctatcgtta tgttgcagtt agctgggggg tggtggtggc
     3661 tgtgacaggc accgtgggca atgtgctcac cctactggcc ttggccatcc agcccaagct
     3721 ccgtacccga ttcaacctgc tcatagccaa cctcacactg gctgatctcc tctactgcac
     3781 gctccttcag cccttctctg tggacaccta cctccacctg cactggcgca ccggtgccac
     3841 cttctgcagg gtatttgggc tcctcctttt tgcctccaat tctgtctcca tcctgaccct
     3901 ctgcctcatc gcactgggac gctacctcct cattgcccac cctaagcttt ttccccaagt
     3961 tttcagtgcc aaggggatag tgctggcact ggtgagcacc tgggttgtgg gcgtggccag
     4021 ctttgctccc ctctggccta tttatatcct ggtacctgta gtctgcacct gcagctttga
     4081 ccgcatccga ggccggcctt acaccaccat cctcatgggc atctactttg tgcttgggct
     4141 cagcagtgtt ggcatcttct attgcctcat ccaccgccag gtcaaacgag cagcacaggc
     4201 actggaccaa tacaagttgc gacaggcaag catccactcc aaccatgtgg ccaggactga
     4261 tgaggccatg cctggtcgtt tccaggagct ggacagcagg ttagcatcag gaggacccag
     4321 tgaggggatt tcatctgagc cagtcagtgc tgccaccacc cagaccctgg aaggggactc
     4381 atcagaagtg ggagaccaga tcaacagcaa gagagctaag cagatggcag agaaaagccc
     4441 tccagaagca tctgccaaag cccagccaat taaaggagcc agaagagctc cggattcttc
     4501 atcggaattt gggaaggtga ctcgaatgtg ttttgctgtg ttcctctgct ttgccctgag
     4561 ctacatcccc ttcttgctgc tcaacattct ggatgccaga gtccaggctc cccgggtggt
     4621 ccacatgctt gctgccaacc tcacctggct caatggttgc atcaaccctg tgctctatgc
     4681 agccatgaac cgccaattcc gccaagcata tggctccatt ttaaaaagag ggccccggag
     4741 tttccatagg ctccattcta gagactacaa agaccatgac ggtgattata aagatcatga
     4801 catcgactac aaggatgacg atgacaagta gcggccgcgg atcccgcccc tctccctccc
     4861 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     4921 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     4981 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     5041 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     5101 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     5161 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     5221 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     5281 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     5341 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     5401 tttgaaaaac acgatgataa gcttgccaca acccacaagg agacgacctt ccatgaccga
     5461 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     5521 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     5581 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5641 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5701 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5761 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5821 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5881 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5941 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     6001 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct agacgcgtct
     6061 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     6121 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     6181 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     6241 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     6301 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     6361 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6421 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     6481 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6541 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6601 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     6661 ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     6721 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     6781 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6841 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6901 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     6961 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     7021 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     7081 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     7141 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     7201 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     7261 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     7321 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7381 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7441 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     7501 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     7561 agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg gcgtaatcat
     7621 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7681 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7741 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7861 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7981 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8041 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8101 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8161 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8221 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8281 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8341 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8401 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8461 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8521 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8581 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8641 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     8701 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     8761 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8821 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8881 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8941 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9001 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9061 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9121 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9181 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9241 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9301 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9361 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9421 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9481 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9541 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     9601 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     9661 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     9721 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     9781 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     9841 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9901 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     9961 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
    10021 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
    10081 actacaaact tagtagt
//
