LOCUS V042664 10258 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042664 VERSION V042664 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..4284 /label="Atf6(NM_001081304)" /note="Atf6(NM_001081304)" /gene="Atf6" CDS 4291..4356 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4400..4611 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4624..4892 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4921..4930 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4951..5613 /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 5683..5736 /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 5737..6336 /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 6337..6925 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6999..7232 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7304..7425 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7465..7600 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7633..7649) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7657..7673 /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(7681..7711) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7726..7747 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8035..8623) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8794..9654) /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(9655..9759) /gene="bla" /label="AmpR promoter" primer_bind 10233..10249 /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 agtcgccttt tagtccggtt 2341 cttcctcatg gaccagatga agactgggag tcgacgttgt ttgctgaact tggctatttc 2401 acagacactg atgatgtgca ctttgatgca gcacatgagg cttatgaaaa taattttgat 2461 catcttaatt ttgatttgga tttgatgcct tgggagtcag acctatggag ccccggcagc 2521 cacttctgct cagacatgaa ggcagagccc cagcctcttt ctccggcttc ctccagttgc 2581 tccatctcct ctcctcggtc cacagactcg tgttcttcaa ctcagcacgt tcctgaggag 2641 ttggatttgt tgtctagttc tcagtccccc ctttccttat atggcgacag ctgtaatagc 2701 ccctcctctg tagagccact gaaggaagag aagcctgtca ctggtcctgg aaacaaaaca 2761 gaacatggac tgactccaaa gaaaaaaatt cagatgagtt caaaaccttc agttcagccc 2821 aagcctttat tacttccagc agcgcccaag actcaaacca atgccagtgt cccagcaaaa 2881 gccatcatca ttcagacact accagccctt atgccactgg caaagcagca gtcgattatc 2941 agcatacagc ctgcgcccac caaaggccag actgttttgc tctctcagcc gactgtggtt 3001 caacttcaga gccctgcggt tctgtcgtct gctcagccgg ttcttgcagt cactggggga 3061 gccgcacagc tacctaacca tgtggtgaat gtgctgccag cccctgtggt gagcagcccg 3121 gtgaatggaa aactttccgt gactaaacct gttctacaaa gtgccaccag aagtatgggt 3181 tcggatatcg ctgtgctgag gagacagcag cggatgataa agaaccgaga gtctgcttgt 3241 cagtcgcgca agaagaagaa agagtatatg ctaggactgg aggccaggct caaggctgcc 3301 ctctcagaga atgagcagct gaagaaggag aatggctccc tgaagcgaca gctggacgag 3361 gtggtgtcag agaaccagag gctcaaagtc ccaagtccaa agcgaagagc tgtctgtgtg 3421 atgatagtat tagcatttat aatgctgaac tatgggccca tgagcatgct ggagcaagaa 3481 tcccgaagag tgaaacctag tgtgagccct gccaatcaga ggaggcatct cttggaattt 3541 tcagcaaaag aagttaaaga cacatcagat ggtgacaacc agaaagacag ttacagctat 3601 gatcactctg tgtccaatga caaagcttta atggtgctaa gtgaagagcc attgctttat 3661 atgcctccac ctccatgtca acccctgatt aacacaacag agtctctcag gttgaaccat 3721 gaacttcgag gctgggttca tagacatgaa gtggaaagga ccaaatctag aagaatgaca 3781 aatagccaac agaaagcccg cattctccag ggtgctctgg aacagggctc taattctcag 3841 ctgatggctg tccagtacac agaaaccact agcatcagta ggaattctgg gagtgagctg 3901 caagtgtatt acgcctcccc tggaagttac caaggcttct ttgacgccat ccgcaggagg 3961 ggagatacgt tttacgttgt ctcatttcga agggatcatc tgctattacc agctaccacc 4021 cacaacaaga ccacaagacc aaaaatgtca attgtattac cagcaataaa cataaatgat 4081 aatgtgatca atgggcagga ctatgaagta atgatgcaga ttgactgtca ggtgatggac 4141 accaggatcc tccacatcaa aagctcctcg gttccccctt atctccggga tcatcagcgg 4201 aaccaaacca gcaccttctt tggttcccct ccaacaacca cagagacgac ccatgtggtc 4261 agcaccatcc ctgagtcgtt gcagggatcc gactacaaag accatgacgg tgattataaa 4321 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 4381 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 4441 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 4501 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4561 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4621 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4681 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4741 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4801 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4861 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4921 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4981 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 5041 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 5101 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 5161 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 5221 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 5281 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 5341 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 5401 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 5461 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 5521 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 5581 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 5641 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 5701 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 5761 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 5821 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5881 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5941 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 6001 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 6061 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 6121 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 6181 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 6241 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 6301 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 6361 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 6421 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 6481 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 6541 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 6601 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 6661 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 6721 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 6781 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6841 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6901 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6961 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 7021 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 7081 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 7141 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 7201 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 7261 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 7321 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 7381 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 7441 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 7501 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 7561 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 7621 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 7681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 7741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8221 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 8341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 8401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 8461 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 8581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 8641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 8701 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 8761 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8821 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8881 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8941 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 9001 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 9061 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 9121 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 9181 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 9241 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 9301 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 9361 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 9421 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 9481 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 9541 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 9601 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 9661 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 9721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9781 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9841 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9901 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9961 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 10021 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 10081 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 10141 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 10201 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //