LOCUS       V030472                 9844 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030472
VERSION     V030472
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             2302..4695
                     /label="USP10(NM_005153)"
                     /note="USP10(NM_005153)"
                     /gene="USP10"
     CDS             4705..4770
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4814..5025
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5038..5306
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5333..5932
                     /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    5939..6527
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6601..6834
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6906..7027
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7046..7181
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7219..7235)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7243..7259
                     /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(7267..7297)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7312..7333
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7621..8209)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8380..9240)
                     /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(9241..9345)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9819..9835
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggccctc cacagcccgc agtatatttt tggagatttt
     2341 agccctgatg aattcaatca attctttgtg actcctcgat cttcagttga gcttcctcca
     2401 tacagtggaa cagttctgtg tggcacacag gctgtggata aactacctga tggacaagaa
     2461 tatcagagaa ttgagtttgg tgtcgatgaa gtcattgaac ccagtgacac tttgccgaga
     2521 acccccagct acagtatttc aagcacactg aaccctcagg cccctgaatt tattctcggt
     2581 tgtacagctt ccaaaataac ccctgatggt atcactaaag aagcaagcta tggctccatc
     2641 gactgccagt acccaggctc tgccctcgct ttggatggaa gttctaatgt ggaggcggaa
     2701 gttttggaaa atgatggtgt ctcaggtggt cttggacaaa gggagcgtaa aaagaagaaa
     2761 aagcggccac ctggatatta cagctatttg aaagatggtg gcgatgatag tatctccaca
     2821 gaagccctgg tcaatggcca tgccaattca gcagtcccga acagtgtcag tgcagaggat
     2881 gcagaattta tgggtgacat gcccccgtca gttacgccca ggacttgtaa cagcccccag
     2941 aactccacag actctgtcag tgacattgtg cctgacagtc ctttccccgg agcactcggc
     3001 agtgacacca ggactgcagg gcagccagag gggggccccg gggctgattt tggtcagtcc
     3061 tgcttccctg cagaggctgg cagagacacc ctgtcaagga cagctggggc tcagccctgc
     3121 gttggtaccg atactactga aaaccttgga gttgctaatg gacaaatact tgaatcctcg
     3181 ggtgagggca cagctaccaa cggggtggag ttgcacacca cggaaagcat agacttggac
     3241 ccaaccaaac ccgagagtgc atcacctcct gctgacggca cgggctctgc atcaggcacc
     3301 cttcctgtca gccagcccaa gtcctgggcc agcctctttc atgattctaa gccctcttcc
     3361 tcctcgccgg tggcctatgt ggaaactaag tattcccctc ccgccatatc tcccctggtt
     3421 tctgaaaagc aggttgaagt caaagaaggg cttgttccgg tttcagagga tcctgtagcc
     3481 ataaagattg cagagttgct ggagaatgta accctaatcc ataaaccagt gtcgttgcaa
     3541 ccccgtgggc tgatcaataa agggaactgg tgctacatta atgctacact gcaggcattg
     3601 gttgcttgcc cgccgatgta ccacctgatg aagttcattc ctctgtattc caaagtgcaa
     3661 aggccttgta cgtcaacacc catgatagac agctttgttc ggctaatgaa tgagttcact
     3721 aatatgccag tacctccaaa accccgacaa gctcttggag ataaaatcgt gagggatatt
     3781 cgccctggag ctgcctttga gcccacatat atttacagac tcctgacagt taacaagtca
     3841 agcctgtctg aaaagggtcg acaagaagat gctgaggaat acttaggctt cattctaaat
     3901 ggacttcatg aggaaatgtt gaacctaaag aagcttctct caccaagtaa tgaaaaactt
     3961 acgatttcca acggccccaa aaaccactcg gtcaatgaag aagagcagga agaacaaggt
     4021 gaaggaagcg aggatgaatg ggaacaagtg ggcccccgga acaagacttc cgtcacccgc
     4081 caggcggatt ttgttcagac tccaatcacc ggcatttttg gtggacacat caggtctgtg
     4141 gtttaccagc agagttcaaa agaatctgcc actttgcagc catttttcac gttgcagttg
     4201 gatatccagt cagacaagat acgcacagtc caggatgcac tggagagctt ggtggcaaga
     4261 gaatctgtcc aaggttatac cacaaaaacc aaacaagagg ttgagataag tcgaagagtg
     4321 actctggaaa aactccctcc tgtcctcgtg ctgcacctga aacgattcgt ttatgagaag
     4381 actggtgggt gccagaagct tatcaaaaat attgaatatc ctgtggactt ggaaattagt
     4441 aaagaactgc tttctccagg ggttaaaaat aagaatttta aatgccaccg aacctatcgg
     4501 ctctttgcag tggtctacca tcacggcaac agtgcgacgg gcggccatta cactacagac
     4561 gtcttccaga tcggtctgaa tggctggctg cgcatcgatg accagacagt caaggtgatc
     4621 aaccagtacc aggtggtgaa accaactgct gaacgcacag cctacctcct gtattaccgc
     4681 cgagtggacc tgctgtaagg atccgactac aaagaccatg acggtgatta taaagatcat
     4741 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     4801 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4861 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     4921 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     4981 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     5041 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     5101 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     5161 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     5221 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     5281 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga
     5341 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     5401 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     5461 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5521 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5581 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5641 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5701 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5761 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5821 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5881 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa
     5941 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     6001 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     6061 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     6121 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     6181 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6241 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     6301 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     6361 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     6421 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6481 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa
     6541 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6601 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     6661 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6721 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6781 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6841 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6901 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7021 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     7081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     7141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     7201 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     7261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8041 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     8101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     9421 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     9481 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     9541 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     9601 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     9661 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     9721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     9781 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9841 gctg
//
