LOCUS       V018244                 9453 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018244
VERSION     V018244
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..3538
                     /label="IDH1(NM_005896)"
                     /note="IDH1(NM_005896)"
                     /gene="IDH1"
     CDS             3539..3562
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        3612..3823
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3836..4104
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4133..4142
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4163..4825
                     /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             4895..4948
                     /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             4949..5548
                     /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    5549..6136
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6210..6443
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6515..6636
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6655..6790
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6828..6844)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6852..6868
                     /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(6876..6906)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6921..6942
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7230..7818)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7989..8849)
                     /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(8850..8954)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9428..9444
                     /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 tagcgaattc gccaccatgt ccaaaaaaat cagtggcggt tctgtggtag agatgcaagg
     2341 agatgaaatg acacgaatca tttgggaatt gattaaagag aaactcattt ttccctacgt
     2401 ggaattggat ctacatagct atgatttagg catagagaat cgtgatgcca ccaacgacca
     2461 agtcaccaag gatgctgcag aagctataaa gaagcataat gttggcgtca aatgtgccac
     2521 tatcactcct gatgagaaga gggttgagga gttcaagttg aaacaaatgt ggaaatcacc
     2581 aaatggcacc atacgaaata ttctgggtgg cacggtcttc agagaagcca ttatctgcaa
     2641 aaatatcccc cggcttgtga gtggatgggt aaaacctatc atcataggtc gtcatgctta
     2701 tggggatcaa tacagagcaa ctgattttgt tgttcctggg cctggaaaag tagagataac
     2761 ctacacacca agtgacggaa cccaaaaggt gacatacctg gtacataact ttgaagaagg
     2821 tggtggtgtt gccatgggga tgtataatca agataagtca attgaagatt ttgcacacag
     2881 ttccttccaa atggctctgt ctaagggttg gcctttgtat ctgagcacca aaaacactat
     2941 tctgaagaaa tatgatgggc gttttaaaga catctttcag gagatatatg acaagcagta
     3001 caagtcccag tttgaagctc aaaagatctg gtatgagcat aggctcatcg acgacatggt
     3061 ggcccaagct atgaaatcag agggaggctt catctgggcc tgtaaaaact atgatggtga
     3121 cgtgcagtcg gactctgtgg cccaagggta tggctctctc ggcatgatga ccagcgtgct
     3181 ggtttgtcca gatggcaaga cagtagaagc agaggctgcc cacgggactg taacccgtca
     3241 ctaccgcatg taccagaaag gacaggagac gtccaccaat cccattgctt ccatttttgc
     3301 ctggaccaga gggttagccc acagagcaaa gcttgataac aataaagagc ttgccttctt
     3361 tgcaaatgct ttggaagaag tctctattga gacaattgag gctggcttca tgaccaagga
     3421 cttggctgct tgcattaaag gtttacccaa tgtgcaacgt tctgactact tgaatacatt
     3481 tgagttcatg gataaacttg gagaaaactt gaagatcaaa ctagctcagg ccaaacttga
     3541 ttacaaggat gacgacgata agtaaggatc cgcggccgca aggatctgcg atcgctccgg
     3601 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     3661 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     3721 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     3781 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc
     3841 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag
     3901 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag
     3961 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact
     4021 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt
     4081 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat
     4141 ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct
     4201 gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat
     4261 gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca
     4321 cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt
     4381 cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg
     4441 cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt
     4501 caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg
     4561 cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt
     4621 cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat
     4681 gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt
     4741 ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc
     4801 cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga
     4861 cggcaccgcc ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg
     4921 cggtgacgtg gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc
     4981 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5041 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5101 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5161 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5221 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5281 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5341 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5401 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5461 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5521 gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag
     5581 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5641 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5701 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5761 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5821 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5881 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5941 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6001 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6061 gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct
     6121 ttggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct
     6181 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata
     6241 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6301 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6361 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6421 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6481 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     6541 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6601 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     6661 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6721 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6781 tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt
     6841 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6901 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6961 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7021 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7081 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7141 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7741 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7801 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7861 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7921 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7981 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8041 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8101 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8161 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8221 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8281 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8341 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8401 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8461 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8521 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8581 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8641 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8701 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8761 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8821 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8881 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8941 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     9001 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     9061 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     9121 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     9181 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     9241 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     9301 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     9361 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     9421 cgacgttgta aaacgacggc cagtgccaag ctg
//
