LOCUS V039563 10213 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039563 VERSION V039563 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..4239 /label="HSPA1A(NM_005345)" /note="HSPA1A(NM_005345)" /gene="HSPA1A" CDS 4246..4311 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4355..4566 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4579..4847 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4876..4885 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4906..5568 /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 5638..5691 /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 5692..6291 /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 6292..6880 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6954..7187 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7259..7380 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7420..7555 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7588..7604) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7612..7628 /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(7636..7666) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7681..7702 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7990..8578) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8749..9609) /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(9610..9714) /gene="bla" /label="AmpR promoter" primer_bind 10188..10204 /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 ccaaagccgc ggcgatcggc 2341 atcgacctgg gcaccaccta ctcctgcgtg ggggtgttcc aacacggcaa ggtggagatc 2401 atcgccaacg accagggcaa ccgcaccacc cccagctacg tggccttcac ggacaccgag 2461 cggctcatcg gggatgcggc caagaaccag gtggcgctga acccgcagaa caccgtgttt 2521 gacgcgaagc ggctgattgg ccgcaagttc ggcgacccgg tggtgcagtc ggacatgaag 2581 cactggcctt tccaggtgat caacgacgga gacaagccca aggtgcaggt gagctacaag 2641 ggggagacca aggcattcta ccccgaggag atctcgtcca tggtgctgac caagatgaag 2701 gagatcgccg aggcgtacct gggctacccg gtgaccaacg cggtgatcac cgtgccggcc 2761 tacttcaacg actcgcagcg ccaggccacc aaggatgcgg gtgtgatcgc ggggctcaac 2821 gtgctgcgga tcatcaacga gcccacggcc gccgccatcg cctacggcct ggacagaacg 2881 ggcaaggggg agcgcaacgt gctcatcttt gacctgggcg ggggcacctt cgacgtgtcc 2941 atcctgacga tcgacgacgg catcttcgag gtgaaggcca cggccgggga cacccacctg 3001 ggtggggagg actttgacaa caggctggtg aaccacttcg tggaggagtt caagagaaaa 3061 cacaagaagg acatcagcca gaacaagcga gccgtgaggc ggctgcgcac cgcctgcgag 3121 agggccaaga ggaccctgtc gtccagcacc caggccagcc tggagatcga ctccctgttt 3181 gagggcatcg acttctacac gtccatcacc agggcgaggt tcgaggagct gtgctccgac 3241 ctgttccgaa gcaccctgga gcccgtggag aaggctctgc gcgacgccaa gctggacaag 3301 gcccagattc acgacctggt cctggtcggg ggctccaccc gcatccccaa ggtgcagaag 3361 ctgctgcagg acttcttcaa cgggcgcgac ctgaacaaga gcatcaaccc cgacgaggct 3421 gtggcctacg gggcggcggt gcaggcggcc atcctgatgg gggacaagtc cgagaacgtg 3481 caggacctgc tgctgctgga cgtggctccc ctgtcgctgg ggctggagac ggccggaggc 3541 gtgatgactg ccctgatcaa gcgcaactcc accatcccca ccaagcagac gcagatcttc 3601 accacctact ccgacaacca acccggggtg ctgatccagg tgtacgaggg cgagagggcc 3661 atgacgaaag acaacaatct gttggggcgc ttcgagctga gcggcatccc tccggccccc 3721 aggggcgtgc cccagatcga ggtgaccttc gacatcgatg ccaacggcat cctgaacgtc 3781 acggccacgg acaagagcac cggcaaggcc aacaagatca ccatcaccaa cgacaagggc 3841 cgcctgagca aggaggagat cgagcgcatg gtgcaggagg cggagaagta caaagcggag 3901 gacgaggtgc agcgcgagag ggtgtcagcc aagaacgccc tggagtccta cgccttcaac 3961 atgaagagcg ccgtggagga tgaggggctc aagggcaaga tcagcgaggc ggacaagaag 4021 aaggtgctgg acaagtgtca agaggtcatc tcgtggctgg acgccaacac cttggccgag 4081 aaggacgagt ttgagcacaa gaggaaggag ctggagcagg tgtgtaaccc catcatcagc 4141 ggactgtacc agggtgccgg tggtcccggg cctgggggct tcggggctca gggtcccaag 4201 ggagggtctg ggtcaggccc caccattgag gaggtagatg gatccgacta caaagaccat 4261 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 4321 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 4381 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 4441 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4501 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4561 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4621 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4681 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4741 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4801 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4861 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4921 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4981 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 5041 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 5101 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 5161 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 5221 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 5281 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 5341 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 5401 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 5461 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 5521 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 5581 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 5641 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5701 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5761 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5821 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5881 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5941 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 6001 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 6061 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 6121 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 6181 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 6241 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 6301 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 6361 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 6421 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6481 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6541 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6601 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6661 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6721 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6781 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6841 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6901 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6961 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 7021 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 7081 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 7141 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 7201 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 7261 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 7321 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 7381 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 7441 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 7501 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 7561 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 7621 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7681 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7741 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7801 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7861 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 8041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 8101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 8161 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 8221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 8281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 8341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 8401 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 8461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 8521 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8641 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8881 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 9001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 9061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 9121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 9181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 9241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 9301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 9361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 9421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 9481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9721 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9781 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9841 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9901 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9961 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 10021 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 10081 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 10141 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 10201 cagtgccaag ctg //