LOCUS       V030314                 9532 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030314
VERSION     V030314
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..3558
                     /label="IDH1(NM_005896)"
                     /note="IDH1(NM_005896)"
                     /gene="IDH1"
     CDS             3565..3630
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3674..3885
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3898..4166
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4195..4204
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4225..4887
                     /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             4957..5010
                     /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             5011..5610
                     /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    5611..6199
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6273..6506
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6578..6699
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6739..6874
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6907..6923)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6931..6947
                     /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(6955..6985)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7000..7021
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7309..7897)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8068..8928)
                     /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(8929..9033)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9507..9523
                     /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 gccaccatgt ccaaaaaaat cagtggcggt
     2341 tctgtggtag agatgcaagg agatgaaatg acacgaatca tttgggaatt gattaaagag
     2401 aaactcattt ttccctacgt ggaattggat ctacatagct atgatttagg catagagaat
     2461 cgtgatgcca ccaacgacca agtcaccaag gatgctgcag aagctataaa gaagcataat
     2521 gttggcgtca aatgtgccac tatcactcct gatgagaaga gggttgagga gttcaagttg
     2581 aaacaaatgt ggaaatcacc aaatggcacc atacgaaata ttctgggtgg cacggtcttc
     2641 agagaagcca ttatctgcaa aaatatcccc cggcttgtga gtggatgggt aaaacctatc
     2701 atcataggtc gtcatgctta tggggatcaa tacagagcaa ctgattttgt tgttcctggg
     2761 cctggaaaag tagagataac ctacacacca agtgacggaa cccaaaaggt gacatacctg
     2821 gtacataact ttgaagaagg tggtggtgtt gccatgggga tgtataatca agataagtca
     2881 attgaagatt ttgcacacag ttccttccaa atggctctgt ctaagggttg gcctttgtat
     2941 ctgagcacca aaaacactat tctgaagaaa tatgatgggc gttttaaaga catctttcag
     3001 gagatatatg acaagcagta caagtcccag tttgaagctc aaaagatctg gtatgagcat
     3061 aggctcatcg acgacatggt ggcccaagct atgaaatcag agggaggctt catctgggcc
     3121 tgtaaaaact atgatggtga cgtgcagtcg gactctgtgg cccaagggta tggctctctc
     3181 ggcatgatga ccagcgtgct ggtttgtcca gatggcaaga cagtagaagc agaggctgcc
     3241 cacgggactg taacccgtca ctaccgcatg taccagaaag gacaggagac gtccaccaat
     3301 cccattgctt ccatttttgc ctggaccaga gggttagccc acagagcaaa gcttgataac
     3361 aataaagagc ttgccttctt tgcaaatgct ttggaagaag tctctattga gacaattgag
     3421 gctggcttca tgaccaagga cttggctgct tgcattaaag gtttacccaa tgtgcaacgt
     3481 tctgactact tgaatacatt tgagttcatg gataaacttg gagaaaactt gaagatcaaa
     3541 ctagctcagg ccaaacttgg atccgactac aaagaccatg acggtgatta taaagatcat
     3601 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     3661 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3721 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3781 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3841 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     3901 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     3961 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     4021 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     4081 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4141 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agacgccacc
     4201 atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat caccggcacc
     4261 ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa gcagggccgc
     4321 atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc
     4381 cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc
     4441 ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac
     4501 ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac
     4561 ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc
     4621 cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct ggtgggcagc
     4681 ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt ggacagccac
     4741 atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc
     4801 ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga gtaccagcac
     4861 gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg
     4921 gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag tcttctaaca
     4981 tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac ggtgcgcctc
     5041 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5101 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5161 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5221 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5281 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5341 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5401 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5461 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5521 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5581 tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa aatttgtgaa
     5641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5761 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5821 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5881 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5941 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6001 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6061 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6121 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6181 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     6241 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     6301 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6361 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6421 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6481 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6541 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6721 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6781 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6841 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga ccaaattcgt
     6901 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7741 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9061 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     9121 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     9181 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     9241 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     9301 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9361 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9421 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9481 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
