LOCUS       V035799                 9523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035799
VERSION     V035799
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..3549
                     /label="PDK4(NM_002612)"
                     /note="PDK4(NM_002612)"
                     /gene="PDK4"
     CDS             3556..3621
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3665..3876
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3889..4157
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4186..4195
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4216..4878
                     /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             4948..5001
                     /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             5002..5601
                     /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    5602..6190
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6264..6497
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6569..6690
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6730..6865
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6898..6914)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6922..6938
                     /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(6946..6976)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6991..7012
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7300..7888)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8059..8919)
                     /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(8920..9024)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9498..9514
                     /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 aggcggcccg cttcgtgctg
     2341 cgcagcgctg gctcgctcaa cggcgccggc ctggtgcccc gagaggtgga gcatttctcg
     2401 cgctacagcc cgtccccgct gtccatgaag cagctactgg actttggttc agaaaatgca
     2461 tgtgaaagaa cttcttttgc atttttgcga caagaattgc ctgtgagact cgccaacatt
     2521 ctgaaggaaa ttgatatcct cccgacccaa ttagtaaata cctcttcagt gcaattggtt
     2581 aaaagctggt atatacagag cctgatggat ttggtggaat tccatgagaa aagcccagat
     2641 gaccagaaag cattatcaga ctttgtagat acactcatca aagttcgaaa tagacaccat
     2701 aatgtagtcc ctacaatggc acaaggaatc atagagtata aagatgcctg tacagttgac
     2761 ccagtcacca atcaaaatct tcaatatttc ttggatcgat tttacatgaa ccgtatttct
     2821 actcggatgc tgatgaacca gcacattctt atatttagtg actcacagac aggaaaccca
     2881 agccacattg gaagcattga tcctaactgt gatgtggtag cagtggtcca agatgccttt
     2941 gagtgttcaa ggatgctctg tgatcagtat tatttatcat ctccagaatt aaagcttaca
     3001 caagtgaatg gaaaatttcc agaccaacca attcacatcg tgtatgttcc ttctcacctc
     3061 catcatatgc tctttgaact atttaagaat gcaatgcggg caacagttga acaccaggaa
     3121 aatcagcctt cccttacacc aatagaggtt attgttgtct tgggaaaaga agaccttacc
     3181 attaagattt cagacagagg aggtggtgtt cccctgagaa ttattgaccg cctctttagt
     3241 tatacatact ccactgcacc aacgcctgtg atggataatt cccggaatgc tcctttggct
     3301 ggttttggtt acggcttgcc aatttctcgt ctgtatgcaa agtactttca aggagatctg
     3361 aatctctact ctttatcagg atatggaaca gatgctatca tctacttaaa ggctttgtct
     3421 tctgagtcta tagaaaaact tccagttttt aacaagtcag ccttcaaaca ttatcagatg
     3481 agctctgagg ctgatgactg gtgtatccca agcagggaac caaagaacct ggcaaaagaa
     3541 gtggccatgg gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3601 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3661 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3721 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3781 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3841 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     3901 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3961 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     4021 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     4081 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     4141 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc
     4201 gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc
     4261 gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac
     4321 aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg
     4381 ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac
     4441 gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg
     4501 ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg
     4561 gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac
     4621 gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc
     4681 accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc
     4741 aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc
     4801 gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag
     4861 acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc
     4921 gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac
     4981 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc
     5041 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5101 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5161 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5221 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5281 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5341 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5401 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5461 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5521 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5581 acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact
     5641 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5701 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5761 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5821 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5881 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5941 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     6001 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     6061 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     6121 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     6181 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     6241 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct
     6301 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6361 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6421 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6481 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6541 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6601 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6661 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6721 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6781 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6841 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc
     6901 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6961 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7021 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7741 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8101 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8161 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8221 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8281 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8341 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8401 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8461 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8521 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8581 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8641 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8701 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8761 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8821 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8881 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8941 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9001 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9061 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     9121 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9181 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9241 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9301 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9361 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9421 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9481 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
