LOCUS       V026682                10001 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026682
VERSION     V026682
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             3548..4648
                     /label="Ccr6(NM_009835)"
                     /note="Ccr6(NM_009835)"
                     /gene="Ccr6"
     CDS             4658..4723
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4751..5337
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5357..5956
                     /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    5970..6558
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6765..7398
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7526..7542)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7550..7566
                     /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(7574..7604)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7619..7640
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7928..8516)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8687..9547)
                     /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(9548..9652)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9700..9834
                     /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 aattccacag agtcctactt tggaacggat gattatgaca acacagagta
     3601 ttattctatt cctccagacc atgggccatg ctccctagaa gaggtcagaa acttcaccaa
     3661 ggtatttgtg ccaattgcct actccttaat atgtgtcttt ggcctcctgg gcaacattat
     3721 ggtggtgatg acctttgcct tctacaagaa agccagatcc atgactgacg tctacctgtt
     3781 gaacatggcc atcacagaca tactctttgt cctcacccta ccgttctggg cagttactca
     3841 tgccaccaac acttgggttt tcagcgatgc actgtgtaaa ctgatgaaag gcacatatgc
     3901 ggtcaacttt aactgtggga tgctgctcct ggcctgtatc agcatggacc ggtacattgc
     3961 catcgtccag gcaaccaaat ctttccgggt acgctccaga acactgacgc acagtaaggt
     4021 catctgtgtg gcagtgtggt tcatctccat catcatctca agccctacat ttatcttcaa
     4081 caagaaatac gagctgcagg atcgtgatgt ctgtgagcca cggtacaggt ctgtctcaga
     4141 gcccatcacg tggaagctgc tgggtatggg actggagctg ttctttgggt tcttcacccc
     4201 tttgctgttt atggtgttct gctatctgtt cattatcaag accttggtgc aggcccagaa
     4261 ctccaagagg cacagagcaa tccgagtcgt gatcgctgtg gttctcgtgt tcctggcttg
     4321 tcagatccct cacaacatgg tcctcctcgt gactgcggtc aacacgggca aagtgggccg
     4381 gagctgcagc accgagaaag tcctcgccta caccaggaac gtggccgagg tcctggcttt
     4441 cctgcattgc tgcctcaacc ccgtgttgta tgcgtttatt ggacagaaat tcagaaacta
     4501 cttcatgaag atcatgaagg atgtgtggtg tatgagaagg aagaataaga tgcctggctt
     4561 cctctgtgcc cgggtttact cggaaagcta catctccagg cagaccagtg agaccgtcga
     4621 aaatgataat gcatcgtcct ttaccatgag cggccgcgac tacaaagacc atgacggtga
     4681 ttataaagat catgacatcg actacaagga tgacgatgac aagtaggcct gcgatatcca
     4741 gcggatcccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     4801 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4861 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4921 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     4981 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5041 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5101 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5161 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     5221 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     5281 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac
     5341 aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5401 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5461 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5521 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5581 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5641 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5701 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5761 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5821 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5881 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5941 caagcccggt gcctagacgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa
     6001 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     6061 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     6121 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6181 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6241 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6301 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6361 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6421 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6481 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6541 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc
     6601 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     6661 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     6721 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca
     6781 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     6841 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     6901 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     6961 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     7021 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     7081 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     7141 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     7201 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     7261 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7321 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     7381 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     7441 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta ccgtcgacct
     7501 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7561 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7621 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7681 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7741 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7801 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7861 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7921 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7981 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8041 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8101 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     8161 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8221 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8281 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8341 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8401 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8461 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8581 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8641 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8701 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8761 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8821 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8881 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8941 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9001 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9061 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9121 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9181 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9241 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9301 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9361 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9421 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9481 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9541 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9601 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9661 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     9721 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9781 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     9841 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     9901 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     9961 agaaattgta tttgttaaat atgtactaca aacttagtag t
//
