LOCUS V026539 8884 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026539 VERSION V026539 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..2910 /label="LCN2(NM_005564)" /note="LCN2(NM_005564)" /gene="LCN2" CDS 2917..2982 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3026..3237 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3250..3518 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3547..3556 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3577..4239 /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 4309..4362 /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 4363..4962 /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 4963..5551 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5625..5858 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5930..6051 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6091..6226 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6259..6275) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6283..6299 /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(6307..6337) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6352..6373 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6661..7249) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7420..8280) /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(8281..8385) /gene="bla" /label="AmpR promoter" primer_bind 8859..8875 /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 ccctaggtct cctgtggctg 2341 ggcctagccc tgttgggggc tctgcatgcc caggcccagg actccacctc agacctgatc 2401 ccagccccac ctctgagcaa ggtccctctg cagcagaact tccaggacaa ccaattccag 2461 gggaagtggt atgtggtagg cctggcaggg aatgcaattc tcagagaaga caaagacccg 2521 caaaagatgt atgccaccat ctatgagctg aaagaagaca agagctacaa tgtcacctcc 2581 gtcctgttta ggaaaaagaa gtgtgactac tggatcagga cttttgttcc aggttgccag 2641 cccggcgagt tcacgctggg caacattaag agttaccctg gattaacgag ttacctcgtc 2701 cgagtggtga gcaccaacta caaccagcat gctatggtgt tcttcaagaa agtttctcaa 2761 aacagggagt acttcaagat caccctctac gggagaacca aggagctgac ttcggaacta 2821 aaggagaact tcatccgctt ctccaaatct ctgggcctcc ctgaaaacca catcgtcttc 2881 cctgtcccaa tcgaccagtg tatcgacggc ggatccgact acaaagacca tgacggtgat 2941 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3001 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3061 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3121 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3181 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3241 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3301 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3361 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3421 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3481 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3541 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 3601 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 3661 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 3721 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 3781 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 3841 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 3901 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 3961 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4021 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4081 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4141 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4201 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4261 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4321 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4381 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4441 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4501 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4561 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4621 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 4681 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 4741 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 4801 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 4861 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 4921 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 4981 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5041 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5101 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5161 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5221 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5281 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5341 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5401 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5461 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc tccgccttcg ccctcagacg 5521 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 5581 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 5641 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 5701 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 5761 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 5821 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 5881 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 5941 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6001 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6061 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6241 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6301 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6361 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6421 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6841 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8401 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8461 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8521 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8581 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8641 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8701 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8761 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8821 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8881 gctg //