LOCUS       V019353                 9990 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019353
VERSION     V019353
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             2827..4716
                     /label="Il4i1(NM_010215)"
                     /note="Il4i1(NM_010215)"
                     /gene="Il4i1"
     CDS             4723..4746
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4778..5277
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5298..5897
                     /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    5911..6499
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6706..7339
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7468..7484)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7492..7508
                     /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(7516..7546)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7561..7582
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7870..8458)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8629..9489)
                     /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(9490..9594)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9642..9776
                     /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 tcgctagcgc taccggactc agatctcgag
     2821 gccaccatgg ctgggctggc cctgcgtctt gtcctggcgg ccaccctcct tggtctggca
     2881 ggctctctgg actggaaggc agcctccagc ttgaacccta ttgagaagtg tatggaggac
     2941 cacgattatg agcagctact caaggtggtg accttgggcc tcaatcggac ttcgaagccc
     3001 cagaaggtgg tagtggttgg tgcaggcgtg gcagggctgg tagcagccaa gatgctcagt
     3061 gatgcaggac acaaggtcac catcctggag gcagataaca ggattggggg ccgtatcttc
     3121 actttccggg atgagaagac aggctggata ggggagcttg gggccatgcg aatgcccagc
     3181 tctcacagga tcttgcacaa gctctgcagg accctgggcc tcaacctgac tcagttcaca
     3241 cagtatgatg agaacacgtg gacggaggtg cataatgtga agctgcgaaa ctatgtggtg
     3301 gagaagatgc cagaaaagct gggctacaac ctgaacaaca gggaaagggg ccattcccca
     3361 gaggacatct accagatggc actcaacaag gccttcaaag acctcaaggc cttgggctgc
     3421 aagaaagcca tgaataagtt caacaagcat acgcttctgg aatacctcct cgaggagggc
     3481 aacctgtccc ggccggctgt gcagctcctg ggagatgtga tgtccgagga gggcttcttc
     3541 tacctcagct ttgcagaagc cttacgtgcg cacgcctgcc tgagcgatag actccggtac
     3601 agccgcatcg taggtggctg ggacctgctt ccgcgagctc tgctgagctc actgtccggg
     3661 gcgctgctac tgaacgcgcc tgtggtgtcg atcactcagg ggaggaacga tgtgcgcgtg
     3721 cacattgcca cctcgcttca cagcgagaag acgctgacag ccgacgtggt gctgctgact
     3781 gccagtggac ccgcgctgca gcgcattacc ttctcgccgc ccttgactcg caagaggcag
     3841 gaggcactgc gcgcgcttca ctacgtagca gccagcaagg tttttctgag tttccgtcgg
     3901 cccttctggc acgaggagca catcgagggc ggccactcca acactgaccg cccatcgcgc
     3961 ctcatattct atcccgcgca gggcgagggc tcactgcttc tggcctccta cacgtggtcg
     4021 gacgctgcag cccccttcgc tggactgagc accgaccaga ccctgcgttt ggtgctccag
     4081 gacgtggcgg ccctgcacgg gcctgtggtg ttccggctgt gggacggcag gggtgtggtc
     4141 aagcgctggg cagaggaccc gcatagccag ggaggcttcg tggtgcagcc gccattgtac
     4201 gggcgcgagg ctgaggacta tgactggtca gcccccttcg gccgcattta cttcgcgggc
     4261 gagcacacag ctctcccgca tggctgggta gagaccgctg tcaagtccgg gttgcgggcc
     4321 gcggtgagaa tcaataataa ctatgggtac ggggaggtcg acccccagat gatggagcat
     4381 gcatatgcag aggccaacta tctggaccag tatcctgaag gggagaggcc cgaggagcag
     4441 caggcgcggg aagaagtcag cccagatgaa caggagccct ctcacaaaca cttgttggtg
     4501 gaaacgagcc ccgaggggca gcaacacgcg tttgtggagg ccattcccga gctgcaggga
     4561 cacgtgttcg tggagactgt cccccaggag aaggggcacg cccaccagaa tatatatcct
     4621 tcggagcatg tacaggtgca tggggaagtc atccctgagt ggcatggtca tgggggatct
     4681 ggcaccccgc aaatgcaccg agtgggggac cactccgaat tcgattacaa ggatgacgac
     4741 gataagtgag gatcccgcga ctctagataa ttctaccggg taggggaggc gcttttccca
     4801 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     4861 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     4921 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     4981 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
     5041 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
     5101 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     5161 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     5221 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
     5281 tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     5341 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     5401 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     5461 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     5521 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     5581 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     5641 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     5701 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     5761 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5821 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5881 gcaagcccgg tgcctgaccg cgtctggaac aatcaacctc tggattacaa aatttgtgaa
     5941 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     6001 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6061 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6121 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6181 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6241 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6301 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6361 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6421 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6481 ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag
     6541 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     6601 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     6661 aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc
     6721 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     6781 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
     6841 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
     6901 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
     6961 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
     7021 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
     7081 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
     7141 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7201 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7261 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7321 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     7381 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt taccgtcgac
     7441 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7501 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7561 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7621 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7681 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8281 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9601 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt
     9661 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9721 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
     9781 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
     9841 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     9901 aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa gaaattgtat
     9961 ttgttaaata tgtactacaa acttagtagt
//
