LOCUS       V041032                 9451 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041032
VERSION     V041032
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"
     CDS             2297..3463
                     /label="Sirt2(NM_022432)"
                     /note="Sirt2(NM_022432)"
                     /gene="Sirt2"
     CDS             3500..3565
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3610..3821
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3834..4102
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4131..4140
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4161..4823
                     /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             4893..4946
                     /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             4947..5546
                     /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    5547..6134
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6208..6441
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6513..6634
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6653..6788
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6826..6842)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6850..6866
                     /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(6874..6904)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6919..6940
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7228..7816)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7987..8847)
                     /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(8848..8952)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9426..9442
                     /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 gtttatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg ccgagccgga cccctctgac cctctggaga cccaggcagg
     2341 gaaggtgcag gaggctcagg attcagactc ggacactgag ggaggagcca ctggtggaga
     2401 ggcagagatg gacttcctga ggaatttatt cacccagacc ctgggcctgg gttcccaaaa
     2461 ggagcgtctt ctagacgagc tgaccctcga aggagtgaca cgctacatgc agagcgagcg
     2521 ctgccgcaag gtcatctgtt tggtgggagc cggaatctcc acgtccgcgg gtatccctga
     2581 cttccgctcc ccgtccactg gcctctatgc aaacctggag aagtaccacc ttccttaccc
     2641 agaggccatc tttgagatca gctacttcaa gaaacatccg gaacccttct ttgcccttgc
     2701 caaggagctc tatcccgggc agttcaagcc aaccatctgc cactacttca tccgcctgct
     2761 gaaggagaag gggctgctgc tgcgctgcta cacgcagaac atagacacgc tggaacgagt
     2821 ggcggggctg gagccccagg acctggtgga ggcccacggc accttctaca catcacactg
     2881 tgtcaacacc tcctgcagaa aagaatacac gatgggctgg atgaaagaga agatcttctc
     2941 agaagcaact cccaggtgtg agcagtgtca gagtgtggta aagcctgata tcgtgttttt
     3001 cggtgagaac cttccatcgc gcttcttctc ctgcatgcag tcagacttct ccaaggtgga
     3061 cctcctcatc atcatgggca cctccctgca ggtgcagccc ttcgcctccc tcatcagcaa
     3121 ggcaccacta gccaccccac ggctgctcat taacaaggaa aagacaggcc agacggaccc
     3181 cttcctgggc atgatgatgg gcctgggagg tggcatggat tttgactcca agaaggctta
     3241 cagggacgtg gcctggctgg gtgactgtga tcaaggctgc ctggctctcg ctgacctcct
     3301 cggatggaag aaggaactgg aagaccttgt ccggagggag catgccaaca tagatgccca
     3361 gtcagggtca caggccccca accccagcac taccatctcc cctggaaagt ccccaccgcc
     3421 tgccaaggag gcggccagga ccaaagagaa agaggaacag cagggcggcg gagggagtgg
     3481 aggaggggga agcggatccg actacaaaga ccatgacggt gattataaag atcatgacat
     3541 cgactacaag gatgacgatg acaagtgagc ggccgcgaag gatctgcgat cgctccggtg
     3601 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     3661 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     3721 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     3781 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     3841 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     3901 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     3961 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     4021 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     4081 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg
     4141 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga
     4201 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga
     4261 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg
     4321 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc
     4381 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg
     4441 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca
     4501 aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca
     4561 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg
     4621 cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc
     4681 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc
     4741 gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct
     4801 tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg
     4861 gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg
     4921 gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca
     4981 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5041 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5101 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5161 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5221 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5281 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5341 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5401 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5461 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5521 gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat
     5581 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5641 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5701 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5761 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5821 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5881 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5941 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6001 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6061 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctcccttt
     6121 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6181 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6241 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6301 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6361 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6421 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6481 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6721 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6781 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9001 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9061 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9361 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9421 acgttgtaaa acgacggcca gtgccaagct g
//
