LOCUS       V027814                 8874 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027814
VERSION     V027814
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..3728
                     /label="TFEB(NM_007162)"
                     /note="TFEB(NM_007162)"
                     /gene="TFEB"
     CDS             3735..3800
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3844..4055
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4068..4336
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4363..4962
                     /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    4969..5557
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5631..5864
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5936..6057
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6076..6211
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6249..6265)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6273..6289
                     /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(6297..6327)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6342..6363
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6651..7239)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7410..8270)
                     /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(8271..8375)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8849..8865
                     /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 atggcgtcac gcatagggtt gcgcatgcag ctcatgcggg
     2341 agcaggcgca gcaggaggag cagcgggagc gcatgcagca acaggctgtc atgcattaca
     2401 tgcagcagca gcagcagcag caacagcagc agctcggagg gccgcccacc ccggccatca
     2461 atacccccgt ccacttccag tcgccaccac ctgtgcctgg ggaggtgttg aaggtgcagt
     2521 cctacctgga gaatcccaca tcctaccatc tgcagcagtc gcagcatcag aaggtgcggg
     2581 agtacctgtc cgagacctat gggaacaagt ttgctgccca catcagccca gcccagggct
     2641 ctccgaaacc cccaccagcc gcctccccag gggtgcgagc tggacacgtg ctgtcctcct
     2701 ccgctggcaa cagtgctccc aatagcccca tggccatgct gcacattggc tccaaccctg
     2761 agagggagtt ggatgatgtc attgacaaca ttatgcgtct ggacgatgtc cttggctaca
     2821 tcaatcctga aatgcagatg cccaacacgc tacccctgtc cagcagccac ctgaatgtgt
     2881 acagcagcga cccccaggtc acagcctccc tggtgggcgt caccagcagc tcctgccctg
     2941 cggacctgac ccagaagcga gagctcacag atgctgagag cagggccctg gccaaggagc
     3001 ggcagaagaa agacaatcac aacttaattg aaaggagacg aaggttcaac atcaatgacc
     3061 gcatcaagga gttgggaatg ctgatcccca aggccaatga cctggacgtg cgctggaaca
     3121 agggcaccat cctcaaggcc tctgtggatt acatccggag gatgcagaag gacctgcaaa
     3181 agtccaggga gctggagaac cactctcgcc gcctggagat gaccaacaag cagctctggc
     3241 tccgtatcca ggagctggag atgcaggctc gagtgcacgg cctccctacc acctccccgt
     3301 ccggcatgaa catggctgag ctggcccagc aggtggtgaa gcaggagctg cctagcgaag
     3361 agggcccagg ggaggccctg atgctggggg ctgaggtccc tgaccctgag ccactgccag
     3421 ctctgccccc gcaagccccg ctgcccctgc ccacccagcc accatcccca ttccatcacc
     3481 tggacttcag ccacagcctg agctttgggg gcagggagga cgagggtccc ccgggctacc
     3541 ccgaacccct ggcgccgggg catggctccc cattccccag cctgtccaag aaggatctgg
     3601 acctcatgct cctggacgac tcactgctac cgctggcctc tgatccactt ctgtccacca
     3661 tgtcccccga ggcctccaag gccagcagcc gccggagcag cttcagcatg gaggagggcg
     3721 atgtgctggg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     3781 acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc
     3841 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     3901 gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     3961 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     4021 ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct
     4081 ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc
     4141 tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc
     4201 tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc
     4261 tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc
     4321 tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc
     4381 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
     4441 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
     4501 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4561 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4621 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4681 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4741 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4801 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4861 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4921 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg
     4981 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5041 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5101 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5161 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5221 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5281 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5341 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5401 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5461 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5521 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac
     5581 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5641 aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga
     5701 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5761 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5821 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     5881 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     5941 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6001 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6061 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6181 cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg
     6241 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6301 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6361 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6421 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6481 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6541 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7081 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7261 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7321 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7381 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7441 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7501 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7561 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7621 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7681 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7741 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7801 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7861 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7921 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7981 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8041 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8101 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8161 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8221 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8281 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8341 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8401 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8461 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8521 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8581 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8641 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     8701 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8761 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8821 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
