LOCUS       V043522                10330 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043522
VERSION     V043522
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             2843..5005
                     /label="LARGE2(NM_152312)"
                     /note="LARGE2(NM_152312)"
                     /gene="LARGE2"
     CDS             5021..5086
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5118..5617
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5638..6237
                     /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    6251..6839
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7046..7679
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7808..7824)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7832..7848
                     /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(7856..7886)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7901..7922
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8210..8798)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8969..9829)
                     /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(9830..9934)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9982..10116
                     /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 ctcaagcttc gaattcgcca ccatgctgcc ccgagggcgc ccccgggcgc tgggggccgc
     2881 cgcgctgttg ctgctgctgc tgctgctcgg attcctcctg ttcggtgggg acctggggtg
     2941 tgagcgccgc gagcctggcg ggcgagcggg ggccccggga tgcttccccg gcccgctcat
     3001 gccacgtgtc cccccagacg ggaggctgcg gagagccgcc gccctcgacg gagacccggg
     3061 ggccggcccc ggggaccaca accgctccga ctgcggcccg cagccgccgc cgccgcccaa
     3121 gtgcgagctc ttgcatgtgg ccatcgtgtg tgcggggcat aactccagcc gagacgtcat
     3181 caccctggtg aagtccatgc tcttctacag gaaaaatcca ctgcacctcc acttggtgac
     3241 tgacgccgtg gccagaaaca tcctggagac gctcttccac acatggatgg tgcctgctgt
     3301 ccgtgtcagc ttttatcatg ccgaccagct caagccccag gtctcctgga tccccaacaa
     3361 gcactactcc ggcctctatg ggctaatgaa gctggtgctg cccagtgcct tgcctgctga
     3421 gctggcccgc gtcattgtcc tggacacgga tgtcaccttc gcctctgaca tctcggagct
     3481 ctgggccctc tttgctcact tttctgacac gcaggcgatc ggtcttgtgg agaaccagag
     3541 tgactggtac ctgggcaacc tctggaagaa ccacaggccc tggcctgcct tgggccgggg
     3601 atttaacaca ggtgtgatcc tgctgcggct ggaccggctc cggcaggctg gctgggagca
     3661 gatgtggagg ctgacagcca ggcgggagct ccttagcctg cctgccacct cactggctga
     3721 ccaggacatc ttcaacgctg tgatcaagga gcacccgggg ctagtgcagc gtctgccttg
     3781 tgtctggaat gtgcagctgt cagatcacac actggccgag cgctgctact ctgaggcgtc
     3841 tgacctcaag gtgatccact ggaactcacc aaagaagctt cgggtgaaga acaagcatgt
     3901 ggaattcttc cgcaatttct acctgacctt cctggagtac gatgggaacc tgctgcggag
     3961 agagctcttt gtgtgcccca gccagccccc acctggtgct gagcagttgc agcaggccct
     4021 ggcacaactg gacgaggaag acccctgctt tgagttccgg cagcagcagc tcactgtgca
     4081 ccgtgtgcat gtcactttcc tgccccatga accgccaccc ccccggcctc acgatgtcac
     4141 ccttgtggcc cagctgtcca tggaccggct gcagatgttg gaagccctgt gcaggcactg
     4201 gcctggcccc atgagcctgg ccttgtacct gacagacgca gaagctcagc agttcctgca
     4261 tttcgtcgag gcctcaccag tgcttgctgc ccggcaggac gtggcctacc atgtggtgta
     4321 ccgtgagggg cccctatacc ccgtcaacca gcttcgcaac gtggccttgg cccaggccct
     4381 cacgccttac gtcttcctca gtgacattga cttcctgcct gcctattctc tctacgacta
     4441 cctcagggcc tccattgagc agctggggct gggcagccgg cgcaaggcag cactggtggt
     4501 gccggcattc gagaccctgc gctaccgctt cagcttcccc cattccaagg tggagctgtt
     4561 ggccttgctg gatgcgggca ctctctacac cttcaggtac cacgagtggc cccgaggcca
     4621 cgcacccaca gactatgccc gctggcggga ggctcaggcc ccgtaccgtg tgcaatgggc
     4681 ggccaactat gaaccctacg tggtggtgcc acgagactgt ccccgctatg atcctcgctt
     4741 tgtgggcttc ggctggaaca aagtggccca cattgtggag ctggatgccc aggaatatga
     4801 gctcctggtg ctgcccgagg ccttcaccat ccatctgccc cacgctccaa gcctggacat
     4861 ctcccgcttc cgctccagcc ccacctatcg tgactgcctc caggccctca aggacgaatt
     4921 ccaccaggac ttgtcccgcc accatggggc tgctgccctc aaatacctcc cagccctgca
     4981 gcagccccag agccctgccc gaggcggtgg aggcggatca gactacaaag accatgacgg
     5041 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagg gatcccgcga
     5101 ctctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     5161 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     5221 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     5281 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     5341 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     5401 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     5461 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     5521 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     5581 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     5641 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5701 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5761 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5821 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5881 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5941 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6001 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6061 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6121 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6181 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg
     6241 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6301 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6361 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6421 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6481 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6541 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6601 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     6661 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6721 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6781 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6841 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca
     6901 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6961 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     7021 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat
     7081 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
     7141 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     7201 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg
     7261 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc
     7321 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat
     7381 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg
     7441 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg
     7501 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     7561 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     7621 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     7681 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     7741 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt
     7801 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7861 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     8041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     8101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     8161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     8221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     8281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     8341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     8401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     8461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     9001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     9061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     9121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     9181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     9241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     9301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     9361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     9421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     9481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9961 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
    10021 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    10081 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
    10141 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
    10201 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    10261 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa
    10321 acttagtagt
//
