LOCUS V035069 8653 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035069 VERSION V035069 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..2679 /label="RPA3(NM_002947)" /note="RPA3(NM_002947)" /gene="RPA3" CDS 2686..2751 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2795..3006 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3019..3287 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3316..3325 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3346..4008 /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 4078..4131 /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 Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 4132..4731 /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 4732..5320 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5394..5627 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5699..5820 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5860..5995 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6028..6044) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6052..6068 /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(6076..6106) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6121..6142 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6430..7018) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7189..8049) /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(8050..8154) /gene="bla" /label="AmpR promoter" primer_bind 8628..8644 /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 tggacatgat ggacttgccc 2341 aggtcgcgca tcaacgccgg catgctagct caattcatcg acaagcctgt ctgcttcgta 2401 gggaggctgg aaaagattca tcccaccgga aaaatgttta ttctttcaga tggagaagga 2461 aaaaatggaa ccatcgagtt gatggaaccc cttgatgaag aaatctctgg aattgtggaa 2521 gtggttggaa gagtaaccgc caaggccacc atcttgtgta catcttatgt ccagtttaaa 2581 gaagatagcc atccttttga tcttggactt tacaatgaag ctgtgaaaat tatccatgac 2641 ttccctcagt tttatccttt agggattgtg caacatgatg gatccgacta caaagaccat 2701 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 2761 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 2821 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 2881 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 2941 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3001 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3061 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3121 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3181 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3241 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 3301 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 3361 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 3421 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 3481 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 3541 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 3601 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 3661 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 3721 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 3781 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 3841 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 3901 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 3961 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4021 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4081 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4141 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4201 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4261 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4321 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4381 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4441 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4501 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4561 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4621 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4681 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 4741 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 4801 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 4861 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 4921 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 4981 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5041 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5101 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5161 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5221 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5281 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5341 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5401 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 5461 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 5521 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 5581 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 5641 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 5701 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 5761 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 5821 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 5881 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 5941 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6001 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6061 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6121 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6181 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6241 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6841 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7681 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7741 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7801 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7861 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7921 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7981 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8041 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8161 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8221 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8281 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8341 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8401 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 8461 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 8521 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 8581 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 8641 cagtgccaag ctg //