LOCUS       V035471                 9232 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035471
VERSION     V035471
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..3258
                     /label="Naa80(NM_019750)"
                     /note="Naa80(NM_019750)"
                     /gene="Naa80"
     CDS             3265..3330
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3374..3585
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3598..3866
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3895..3904
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3925..4587
                     /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             4657..4710
                     /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             4711..5310
                     /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    5311..5899
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5973..6206
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6278..6399
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6439..6574
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6607..6623)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6631..6647
                     /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(6655..6685)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6700..6721
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7009..7597)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7768..8628)
                     /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(8629..8733)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9207..9223
                     /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 gccaccatgg agctgatcct gagtaccagc
     2341 ccagcaaagc tgactctaga tcctgcacgc cagccagagt tgaccctgag attcaacctg
     2401 tccaagctaa ccctggatcc tgcacgccag ccagagctgt cgctgagtcc taggctagct
     2461 gagctgaccc tggatcccac atgccaccca gagatgagcc tcagtcctgg cccagctgag
     2521 cttaccctgg atcctcaaca ccaggcaaag gagctcccag ttcccaagct gcctgaatta
     2581 atcctggagc ctgtacactg ccgacccgag ctcatgagtg cctgtgctga tctcatcaat
     2641 gaccagtggc cccgcagtcg tgcctcccgt ctccactccc tgggccagtc ctcagatgcc
     2701 ttccccctct gcctgatgct gctgagcccc caacccacac ctggagcagc ccctgttgtg
     2761 gtgggccatg cccgcttatc acgggtacta gaccaacccc acagcctctt agtagagaca
     2821 gtggtggtag cccggcctct gaggggccgt ggctttggtc gccgcctcat ggaaggcttg
     2881 gaggcctttg ccagagcccg gggtttccga cggctacacc tcaccactca tgaccagctg
     2941 tacttctatg cccatctggg ctaccaactg ggtgagcctg tgcagggttt ggccttcacc
     3001 aaccgtcgac tgtccaccac tgtcctacgt gccttctcta agccaccctg cccccagcca
     3061 ccctgcaagg agcctatcct agctgcccaa gctgtcccaa ggagctccaa gggaccccca
     3121 ttgccaccac ctcctcccct accccagtct ctgaccgcct caccgcctcc ttcacccgag
     3181 ccccttcctc aaagcccact agagacatgc tatcgagatc tgaaggggtg ccctatattc
     3241 tggatggaaa aagacatcgg atccgactac aaagaccatg acggtgatta taaagatcat
     3301 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     3361 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3421 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3481 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3541 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     3601 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     3661 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     3721 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     3781 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     3841 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agacgccacc
     3901 atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat caccggcacc
     3961 ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa gcagggccgc
     4021 atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc
     4081 cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc
     4141 ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac
     4201 ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac
     4261 ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc
     4321 cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct ggtgggcagc
     4381 ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt ggacagccac
     4441 atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc
     4501 ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga gtaccagcac
     4561 gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg
     4621 gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag tcttctaaca
     4681 tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac ggtgcgcctc
     4741 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4801 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4861 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4921 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4981 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5041 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5101 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5161 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5221 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5281 tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa aatttgtgaa
     5341 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5401 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5461 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5521 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5581 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5641 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     5701 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     5761 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5821 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5881 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     5941 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     6001 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6061 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6121 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6181 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6241 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6301 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6361 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6421 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6481 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6541 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga ccaaattcgt
     6601 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7441 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8761 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8821 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8881 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8941 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     9001 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9061 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9121 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9181 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
