LOCUS       V022574                 9270 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022574
VERSION     V022574
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             2303..3352
                     /label="ATP6AP2(NM_005765)"
                     /note="ATP6AP2(NM_005765)"
                     /gene="ATP6AP2"
     CDS             3353..3379
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3429..3640
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3653..3921
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3950..3959
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3980..4642
                     /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             4712..4765
                     /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             4766..5365
                     /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    5366..5953
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6027..6260
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6332..6453
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6472..6607
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6645..6661)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6669..6685
                     /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(6693..6723)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6738..6759
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7047..7635)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7806..8666)
                     /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(8667..8771)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9245..9261
                     /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 gaattcgcca ccatggctgt gtttgtcgtg ctcctggcgt tggtggcggg
     2341 tgttttgggg aacgagttta gtatattaaa atcaccaggg tctgttgttt tccgaaatgg
     2401 aaattggcct ataccaggag agcggatccc agacgtggct gcattgtcca tgggcttctc
     2461 tgtgaaagaa gacctttctt ggccaggact cgcagtgggt aacctgtttc atcgtcctcg
     2521 ggctaccgtc atggtgatgg tgaagggagt gaacaaactg gctctacccc caggcagtgt
     2581 catttcgtac cctttggaga atgcagttcc ttttagtctt gacagtgttg caaattccat
     2641 tcactcctta ttttctgagg aaactcctgt tgttttgcag ttggctccca gtgaggaaag
     2701 agtgtatatg gtagggaagg caaactcagt gtttgaagac ctttcagtca ccttgcgcca
     2761 gctccgtaat cgcctgtttc aagaaaactc tgttctcagt tcactccccc tcaattctct
     2821 gagtaggaac aatgaagttg acctgctctt tctttctgaa ctgcaagtgc tacatgatat
     2881 ttcaagcttg ctgtctcgtc ataagcatct agccaaggat cattctcctg atttatattc
     2941 actggagctg gcaggtttgg atgaaattgg gaagcgttat ggggaagact ctgaacaatt
     3001 cagagatgct tctaagatcc ttgttgacgc tctgcaaaag tttgcagatg acatgtacag
     3061 tctttatggt gggaatgcag tggtagagtt agtcactgtc aagtcatttg acacctccct
     3121 cattaggaag acaaggacta tccttgaggc aaaacaagcg aagaacccag caagtcccta
     3181 taaccttgca tataagtata attttgaata ttccgtggtt ttcaacatgg tactttggat
     3241 aatgatcgcc ttggccttgg ctgtgattat cacctcttac aatatttgga acatggatcc
     3301 tggatatgat agcatcattt ataggatgac aaaccagaag attcgaatgg attacccata
     3361 cgacgtacct gattatgcat aggatccgcg gccgcgaagg atctgcgatc gctccggtgc
     3421 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3481 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3541 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3601 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     3661 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     3721 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     3781 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     3841 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     3901 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgga
     3961 gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa
     4021 cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg gccgcatgac
     4081 caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt
     4141 gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct
     4201 gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg
     4261 cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa
     4321 ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag
     4381 caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc
     4441 ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca
     4501 cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg
     4561 ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt
     4621 caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg ccgtggacgg
     4681 caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc taacatgcgg
     4741 tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac
     4801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5341 catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt
     5401 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5461 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5521 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5581 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5641 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5701 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5761 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5821 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5881 ggctctgcgg cctcttccgc gtctccgcct tcgccctcag acgagtcgga tctccctttg
     5941 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6001 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     6061 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6121 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6181 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6241 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6301 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6361 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6421 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6481 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6541 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6601 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     6661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8821 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     8881 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     8941 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     9001 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     9061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     9121 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9181 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9241 cgttgtaaaa cgacggccag tgccaagctg
//
