LOCUS       V035276                 9367 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035276
VERSION     V035276
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3393
                     /label="AGTR1(NM_000685)"
                     /note="AGTR1(NM_000685)"
                     /gene="AGTR1"
     CDS             3400..3465
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3509..3720
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3733..4001
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4030..4039
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4060..4722
                     /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             4792..4845
                     /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             4846..5445
                     /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    5446..6034
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6108..6341
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6413..6534
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6574..6709
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6742..6758)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6766..6782
                     /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(6790..6820)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6835..6856
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7144..7732)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7903..8763)
                     /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(8764..8868)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9342..9358
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga ttctcaactc ttctactgaa
     2341 gatggtatta aaagaatcca agatgattgt cccaaagctg gaaggcataa ttacatattt
     2401 gtcatgattc ctactttata cagtatcatc tttgtggtgg gaatatttgg aaacagcttg
     2461 gtggtgatag tcatttactt ttatatgaag ctgaagactg tggccagtgt ttttcttttg
     2521 aatttagcac tggctgactt atgcttttta ctgactttgc cactatgggc tgtctacaca
     2581 gctatggaat accgctggcc ctttggcaat tacctatgta agattgcttc agccagcgtc
     2641 agtttcaacc tgtacgctag tgtgtttcta ctcacgtgtc tcagcattga tcgatacctg
     2701 gctattgttc acccaatgaa gtcccgcctt cgacgcacaa tgcttgtagc caaagtcacc
     2761 tgcatcatca tttggctgct ggcaggcttg gccagtttgc cagctataat ccatcgaaat
     2821 gtatttttca ttgagaacac caatattaca gtttgtgctt tccattatga gtcccaaaat
     2881 tcaaccctcc cgatagggct gggcctgacc aaaaatatac tgggtttcct gtttcctttt
     2941 ctgatcattc ttacaagtta tactcttatt tggaaggccc taaagaaggc ttatgaaatt
     3001 cagaagaaca aaccaagaaa tgatgatatt tttaagataa ttatggcaat tgtgcttttc
     3061 tttttctttt cctggattcc ccaccaaata ttcacttttc tggatgtatt gattcaacta
     3121 ggcatcatac gtgactgtag aattgcagat attgtggaca cggccatgcc tatcaccatt
     3181 tgtatagctt attttaacaa ttgcctgaat cctctttttt atggctttct ggggaaaaaa
     3241 tttaaaagat attttctcca gcttctaaaa tatattcccc caaaagccaa atcccactca
     3301 aacctttcaa caaaaatgag cacgctttcc taccgcccct cagataatgt aagctcatcc
     3361 accaagaagc ctgcaccatg ttttgaggtt gagggatccg actacaaaga ccatgacggt
     3421 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg
     3481 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     3541 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac
     3601 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     3661 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     3721 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
     3781 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
     3841 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct
     3901 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
     3961 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
     4021 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc
     4081 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc
     4141 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc
     4201 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc
     4261 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc
     4321 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc
     4381 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc
     4441 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac
     4501 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc
     4561 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg
     4621 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc
     4681 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg
     4741 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga
     4801 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag
     4861 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4921 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4981 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5041 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5101 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5161 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5221 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5281 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5341 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5401 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat
     5461 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5521 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5581 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5641 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5701 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5761 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5821 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5881 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5941 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6001 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     6061 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     6121 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca
     6181 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6241 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6301 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     6361 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     6421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6541 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6601 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6661 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6721 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6781 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6841 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7561 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8821 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8881 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8941 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     9001 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     9061 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9121 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9181 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9241 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9301 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9361 caagctg
//
