LOCUS       V021664                 8613 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021664
VERSION     V021664
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2301..3467
                     /label="CBX8(NM_020649)"
                     /note="CBX8(NM_020649)"
                     /gene="CBX8"
     CDS             3474..3539
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3583..3794
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3807..4075
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4102..4701
                     /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    4708..5296
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5370..5603
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5675..5796
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5815..5950
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5988..6004)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6012..6028
                     /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(6036..6066)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6081..6102
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6390..6978)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7149..8009)
                     /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(8010..8114)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8588..8604
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atggagcttt cagcggtggg ggagcgggtg ttcgcggccg
     2341 aagccctcct gaagcggcgc atacggaaag gacgcatgga atacctcgtg aaatggaagg
     2401 gatggtcgca gaagtacagc acatgggaac cggaggaaaa catcctggat gctcgcttgc
     2461 tcgcagcctt tgaggaaagg gaaagagaga tggagctcta tggccccaaa aagcgtggac
     2521 ccaagcccaa aaccttcctc ctcaaagcgc aggccaaggc aaaggccaaa acttacgagt
     2581 ttcgaagtga ctcagccagg ggcatccgga tcccctaccc tggccgctcg ccccaggacc
     2641 tggcctccac ttcccgggcc cgggagggcc ttcgaaacat gggtttgtcc ccgccagcga
     2701 gcagcaccag caccagcagc acctgccgcg cagaggcccc tcgggaccgg gaccgagacc
     2761 gggataggga ccgggagcgg gatcgagaaa gggagaggga gcgagagagg gagcgggaac
     2821 gtgagaggga acgagagcgg ggtaccagca gagtggatga caagcccagc tcaccggggg
     2881 acagctcgaa gaagcgaggc cccaagcccc ggaaggagct cccggacccc tcacagaggc
     2941 ccttaggcga acccagcgcc ggcctcggag agtacctcaa gggcaggaag ctggacgaca
     3001 ccccttccgg ggcaggaaag tttccagccg gccacagtgt gatccagctg gcccgaagac
     3061 aggactcgga cctggtgcag tgtggtgtga ccagccctag ctcagctgag gccacgggca
     3121 aactggctgt ggacaccttc ccggccaggg tgataaagca cagggctgcc ttcctggagg
     3181 ccaaaggcca gggtgcccta gatcccaatg gcacccgggt ccgacatggc tcaggccccc
     3241 ccagctctgg ggggggcctg taccgggaca tgggggccca ggggggaagg ccctccctca
     3301 tcgccaggat ccctgtggcc agaatcctgg gggacccgga ggaagagtcc tggagcccct
     3361 ccctgactaa cctggagaag gtggtggtca cggacgtgac ctcaaacttt ttgaccgtca
     3421 ccattaagga aagtaacacg gaccaaggct tttttaaaga gaaaagagga tccgactaca
     3481 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3541 agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc
     3601 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt
     3661 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     3721 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     3781 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc
     3841 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg
     3901 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt
     3961 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc
     4021 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc
     4081 tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4141 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4201 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4261 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4321 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4381 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4441 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4501 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4561 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4621 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4681 acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     4741 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4801 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4861 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     4921 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     4981 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5041 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5101 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     5161 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     5221 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5281 tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct
     5341 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata
     5401 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     5461 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     5521 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     5581 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     5641 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     5701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5761 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     5821 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5881 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5941 tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt
     6001 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6061 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6121 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6181 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6241 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6301 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6361 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6421 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6481 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6541 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6601 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6661 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6721 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6781 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6841 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6901 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6961 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7021 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7081 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7141 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7201 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7261 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7321 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7381 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7441 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7501 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7561 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7621 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7681 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7741 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7801 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7861 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7921 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7981 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8041 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8101 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     8161 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     8221 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     8281 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     8341 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     8401 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     8461 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     8521 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     8581 cgacgttgta aaacgacggc cagtgccaag ctg
//
