LOCUS       V045086                 9214 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045086
VERSION     V045086
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      2279..2288
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2288..2311
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2324..3322
                     /label="Sirt4(NM_133760)"
                     /note="Sirt4(NM_133760)"
                     /gene="Sirt4"
     promoter        3373..3584
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3597..3865
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3894..3903
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3924..4586
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4656..4709
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4710..5309
                     /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    5310..5897
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5971..6204
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6276..6397
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6416..6551
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6589..6605)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6613..6629
                     /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(6637..6667)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6682..6703
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6991..7579)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7750..8610)
                     /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(8611..8715)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9189..9205
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 caccatggat tacaaggatg acgacgataa ggctagcgaa ttcatgagcg gattgacttt
     2341 caggccgaca aagggccgtt ggatcaccca cctcagccgg ccgcgttctt gtggaccctc
     2401 ggggttattt gtgccgccca gccctccttt ggaccctgaa aagatcaaag agttacagcg
     2461 cttcattagc ctttccaaga aactcctcgt gatgacaggc gcggggatct ccaccgagtc
     2521 cggcatccca gactacaggt cagaaaaggt gggactttac gcccgcactg accggagacc
     2581 catccagcac attgatttcg tccgcagtgc tccggtccgc cagcggtact gggcccgaaa
     2641 ctttgtgggc tggcctcaat tctcctctca ccaacccaac ccagcacact gggctctgag
     2701 caactgggag agactgggga agctgcactg gttggtgact cagaacgtgg acgctttgca
     2761 ctccaaagca gggagtcagc ggctgacgga gctccacgga tgcatgcaca gagtcctgtg
     2821 cctgaactgt ggggagcaga ctgcccgcag ggtgctgcag gaacgcttcc aagccctgaa
     2881 ccccagctgg agcgccgagg cgcagggcgt ggctcccgac ggcgacgtgt tcctcactga
     2941 ggagcaggtc cggagctttc aggtcccgtg ctgtgatcga tgcggcggcc ctctgaaacc
     3001 ggacgtcgtt ttctttgggg acacggtgaa cccagacaag gttgactttg tgcaccggcg
     3061 tgtgaaagag gcggactccc tactggtggt gggatcatcc ctgcaggtgt actctggtta
     3121 caggttcatc ctcaccgccc gcgagcaaaa gctcccaata gccattctga atatcggccc
     3181 cacccggtct gacgatttgg cttgcctgaa gctggattcc cgctgtggag agttgctgcc
     3241 tttaatagac ccgcggagac agcactctga tgtccaaagg ctggaaatga actttcctct
     3301 gagttccgct gctcaagatc cctaaggatc cgcggccgcg aaggatctgc gatcgctccg
     3361 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     3421 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     3481 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     3541 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg
     3601 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga
     3661 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa
     3721 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac
     3781 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt
     3841 tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta gacgccacca
     3901 tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc
     3961 tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag cagggccgca
     4021 tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc
     4081 acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct
     4141 tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg
     4201 gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact
     4261 tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc
     4321 gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg gtgggcagct
     4381 tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg gacagccaca
     4441 tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct
     4501 tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag taccagcacg
     4561 ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg
     4621 acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt cttctaacat
     4681 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg
     4741 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4801 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4861 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4921 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4981 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5041 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5101 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5161 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5221 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5281 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa
     5341 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5401 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5461 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5521 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5581 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5641 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5701 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5761 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5821 tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc
     5881 tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5941 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6001 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6061 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6121 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6181 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6241 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6301 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6421 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6481 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6541 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6601 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6661 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6721 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6781 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6841 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6901 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8341 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8761 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8821 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8881 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8941 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9001 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9061 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9121 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9181 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
