LOCUS       V023327                10000 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023327
VERSION     V023327
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..4696
                     /label="FBXO24(NM_012172)"
                     /note="FBXO24(NM_012172)"
                     /gene="FBXO24"
     CDS             4703..4768
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4788..5287
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5308..5907
                     /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    5921..6509
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6716..7349
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7478..7494)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7502..7518
                     /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(7526..7556)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7571..7592
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7880..8468)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8639..9499)
                     /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(9500..9604)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9652..9786
                     /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 ccatggtgtg ggaaagccag caggaacggg ggggccaagg
     2881 gcctaggaga caggagggca ggagcaggga gggggcaatc aggatggtca ggggaagcca
     2941 gagacggaag ctgcaaaaat tcaccaggct gtgtggacgg gaggaggggc tccgggtgaa
     3001 gagaagctgc ccttcttgtg gctcggagct tggggttgaa gagaagaggg ggaaaggaaa
     3061 tccgatttcc atccagttgt tccccccaga gctggtggag catatcatct cattcctccc
     3121 agtcagagac cttgttgccc tcggccagac ctgccgctac ttccacgaag tgtgcgatgg
     3181 ggaaggcgtg tggagacgca tctgtcgcag actcagtccg cgcctccaag atcagggttc
     3241 tggagtccgg ccctggaaga gagctgccat tctgaactac acgaagggcc tgtatttcca
     3301 ggcatttgga ggccgccgcc gatgtctcag caagagcgtg gcccccttgc tagcccacgg
     3361 ctaccgccgc ttcttgccca ccaaggatca cgtcttcatt cttgactacg tggggaccct
     3421 cttcttcctc aaaaatgccc tggtctccac cctcggccag atgcagtgga agcgggcctg
     3481 tcgctatgtt gtgttgtgtc gtggagccaa ggattttgcc tcggacccaa ggtgtgacac
     3541 agtttaccgt aaatacctct acgtcttggc cactcgggag ccgcaggaag tggtgggtac
     3601 caccagcagc cgggcctgtg actgtgttga ggtctatctg cagtctagtg ggcagcgggt
     3661 cttcaagatg acattccacc actcaatgac cttcaagcag atcgtgctgg ttggtcagga
     3721 gacccagcgg gctctactgc tcctcacaga ggaaggaaag atctactctt tggtagtgaa
     3781 tgagacccag cttgaccagc cacgctccta cacggttcag ctggccctga ggaaggtgtc
     3841 ccactacctg cctcacctgc gcgtggcctg catgacttcc aaccagagca gcaccctcta
     3901 cgtcacagac caggggggag tgtattttga ggtgcatacc ccaggggtgt atcgcgatct
     3961 ctttgggacc cttcaagcct ttgaccccct ggaccagcag atgccgcttg ctctctcact
     4021 gcctgccaag atcctattct gtgctcttgg ctacaaccac cttggcctgg tggatgaatt
     4081 tggccgaatc ttcatgcaag gaaataacag atacgggcag ctaggaacag gggacaaaat
     4141 ggaccgaggg gaacccacac aggtttgtta cctgcagcgg cccatcaccc tgtggtgcgg
     4201 cctcaaccac tccctggtgc tgagccagag ctcagagttc agcaaggagc tgctgggctg
     4261 cggctgtggg gctgggggcc gcctcccagg ctggcccaag gggagtgcct ccttcgtcaa
     4321 gctccaagtc aaggtccctc tgtgtgcctg tgccctctgt gccaccaggg agtgcctata
     4381 catcctgtcc agccacgaca ttgagcagca cgccccctat cgccacctgc cagccagcag
     4441 ggtggtgggg actcctgagc ccagcctggg ggccagagca ccccaggacc ccggggggat
     4501 ggcccaggcc tgcgaggagt acctcagcca gatccacagt tgccaaacgt tgcaggaccg
     4561 cacggagaag atgaaggaga tcgtagggtg gatgcccctg atggccgcac agaaggactt
     4621 cttctgggag gccctggaca tgctgcagag ggctgaagga ggcgggggtg gtgtagggcc
     4681 cccagcccct gagaccggat ccgactacaa agaccatgac ggtgattata aagatcatga
     4741 catcgactac aaggatgacg atgacaagta gtctagataa ttctaccggg taggggaggc
     4801 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     4861 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     4921 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     4981 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     5041 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     5101 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     5161 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     5221 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     5281 cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc
     5341 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5401 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5461 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5521 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     5581 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     5641 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     5701 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     5761 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     5821 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     5881 tgcatgaccc gcaagcccgg tgcctgaccg cgtctggaac aatcaacctc tggattacaa
     5941 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6001 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6061 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6121 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6181 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6241 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6301 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     6361 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6421 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6481 tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa
     6541 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
     6601 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
     6661 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa
     6721 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
     6781 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc
     6841 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa
     6901 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
     6961 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg
     7021 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg
     7081 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc
     7141 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
     7201 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7261 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7321 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     7381 tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt
     7441 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7501 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7561 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7621 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7681 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7741 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7801 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8041 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     8101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8281 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     8341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8401 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8461 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8521 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8581 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8641 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8701 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8761 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8821 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8881 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8941 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     9001 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     9061 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9121 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9181 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9241 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9301 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9361 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9421 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9481 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9541 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9601 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt
     9661 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     9721 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     9781 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
     9841 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
     9901 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa
     9961 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
