LOCUS       V035054                 8563 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035054
VERSION     V035054
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..2589
                     /label="Ccl5(NM_013653)"
                     /note="Ccl5(NM_013653)"
                     /gene="Ccl5"
     CDS             2596..2661
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        2705..2916
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             2929..3197
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3226..3235
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3256..3918
                     /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             3988..4041
                     /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             4042..4641
                     /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    4642..5230
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5304..5537
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5609..5730
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5770..5905
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5938..5954)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5962..5978
                     /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(5986..6016)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6031..6052
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6340..6928)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7099..7959)
                     /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(7960..8064)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8538..8554
                     /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 agatctctgc agctgccctc
     2341 accatcatcc tcactgcagc cgccctctgc acccccgcac ctgcctcacc atatggctcg
     2401 gacaccactc cctgctgctt tgcctacctc tccctcgcgc tgcctcgtgc ccacgtcaag
     2461 gagtatttct acaccagcag caagtgctcc aatcttgcag tcgtgtttgt cactcgaagg
     2521 aaccgccaag tgtgtgccaa cccagagaag aagtgggttc aagaatacat caactatttg
     2581 gagatgagcg gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     2641 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     2701 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     2761 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     2821 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     2881 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     2941 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3001 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     3061 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     3121 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     3181 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc
     3241 gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc
     3301 gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac
     3361 aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg
     3421 ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac
     3481 gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg
     3541 ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg
     3601 gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac
     3661 gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc
     3721 accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc
     3781 aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc
     3841 gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag
     3901 acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc
     3961 gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac
     4021 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4081 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4141 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4201 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4261 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4321 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4381 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4441 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4501 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4561 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4621 acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact
     4681 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4741 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4801 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4861 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4921 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4981 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5041 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5101 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5161 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5221 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5281 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct
     5341 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5401 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5461 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5521 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     5581 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     5641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5701 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5761 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5821 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5881 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc
     5941 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6001 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6061 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6121 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6181 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6241 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6301 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6361 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6421 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6481 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6541 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6601 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6661 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6721 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6781 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6841 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6901 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6961 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7021 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7081 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7141 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7201 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7261 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7321 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7381 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7441 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7501 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7561 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7621 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7681 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7741 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7801 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7861 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7921 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7981 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8041 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8101 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8161 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8221 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8281 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8341 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8401 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     8461 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     8521 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
