LOCUS V026542 9538 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026542 VERSION V026542 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..3564 /label="Calr(NM_007591)" /note="Calr(NM_007591)" /gene="Calr" CDS 3571..3636 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3680..3891 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3904..4172 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4201..4210 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4231..4893 /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 4963..5016 /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 5017..5616 /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 5617..6205 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6279..6512 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6584..6705 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6745..6880 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6913..6929) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6937..6953 /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(6961..6991) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7006..7027 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7315..7903) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8074..8934) /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(8935..9039) /gene="bla" /label="AmpR promoter" primer_bind 9513..9529 /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 gccaccatgc tcctttcggt gccgctcctg 2341 cttggcctcc tcggcctggc cgccgcagac cctgccatct atttcaaaga gcagttcttg 2401 gacggagatg cctggaccaa ccgctgggtc gaatccaaac ataagtccga ttttggcaaa 2461 tttgtcctca gttctggcaa attttacggg gacctggaga aggataaagg gctgcagaca 2521 agccaagatg cccgatttta cgcactgtcc gccaaattcg aacccttcag caataagggc 2581 cagacactgg tggtacagtt cacggtgaag catgagcaga atatcgactg tgggggcggc 2641 tacgtgaagc tgtttccgag tggtttggac cagaaggaca tgcatggaga ctcagaatat 2701 aacatcatgt ttggtccgga catctgcggt cctggcacca agaaggttca tgtcatcttt 2761 aactacaagg gcaagaatgt gctgatcaac aaggatatcc ggtgtaagga tgatgaattc 2821 acacacctat acacactgat tgtgcggcca gacaacacct atgaggtgaa aattgacaac 2881 agccaggtgg agtcaggctc cttggaggat gattgggact ttctgccacc caagaagata 2941 aaggaccctg atgctgccaa gccggaagac tgggatgaac gagccaagat cgatgacccc 3001 acagattcca agcctgagga ctgggacaag ccagagcaca tccctgaccc tgatgctaag 3061 aagcctgagg actgggatga agagatggat ggagagtggg aaccaccagt gattcaaaat 3121 cctgaataca agggcgagtg gaaaccacgt caaattgaca acccagatta caagggtacc 3181 tggatacacc cagaaattga caaccctgaa tactcccccg atgcaaatat ctatgcctat 3241 gatagttttg ctgtactggg cctagatctc tggcaggtca agtccgggac aatctttgac 3301 aatttcctca tcaccaatga tgaggcctat gcagaggagt ttggcaatga gacgtggggt 3361 gttaccaagg ctgcagagaa gcagatgaag gacaagcagg atgaggagca gaggcttaag 3421 gaagaagaag aggacaagaa gcgtaaagag gaagaagaag ctgaggataa agaggatgat 3481 gatgacagag atgaagatga ggacgaagaa gatgagaagg aggaagatga ggaagaatcc 3541 cctggccaag ccaaggatga gctgggatcc gactacaaag accatgacgg tgattataaa 3601 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3661 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3721 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3781 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3841 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3901 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3961 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4021 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4081 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4141 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4201 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4261 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4321 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4381 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4441 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4501 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4561 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4621 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4681 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4741 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4801 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4861 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4921 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4981 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 5041 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 5101 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5161 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5221 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5281 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5341 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5401 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5461 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5521 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5581 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5641 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5701 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5761 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5821 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5881 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5941 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 6001 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 6061 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6121 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6181 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6241 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6301 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6361 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6421 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6481 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6541 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6601 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6661 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6721 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6841 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6901 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6961 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 7021 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7741 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 8041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 9001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9061 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9121 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9181 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9241 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9301 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9361 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9421 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9481 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //