LOCUS V044162 9244 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044162 VERSION V044162 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..3270 /label="APEX1(NM_001641)" /note="APEX1(NM_001641)" /gene="APEX1" CDS 3277..3342 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3386..3597 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3610..3878 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3907..3916 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3937..4599 /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 4669..4722 /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 4723..5322 /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 5323..5911 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5985..6218 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6290..6411 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6451..6586 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6619..6635) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6643..6659 /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(6667..6697) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6712..6733 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7021..7609) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7780..8640) /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(8641..8745) /gene="bla" /label="AmpR promoter" primer_bind 9219..9235 /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 cgaagcgtgg gaaaaaggga 2341 gcggtggcgg aagacgggga tgagctcagg acagagccag aggccaagaa gagtaagacg 2401 gccgcaaaga aaaatgacaa agaggcagca ggagagggcc cagccctgta tgaggacccc 2461 ccagatcaga aaacctcacc cagtggcaaa cctgccacac tcaagatctg ctcttggaat 2521 gtggatgggc ttcgagcctg gattaagaag aaaggattag attgggtaaa ggaagaagcc 2581 ccagatatac tgtgccttca agagaccaaa tgttcagaga acaaactacc agctgaactt 2641 caggagctgc ctggactctc tcatcaatac tggtcagctc cttcggacaa ggaagggtac 2701 agtggcgtgg gcctgctttc ccgccagtgc ccactcaaag tttcttacgg cataggcgat 2761 gaggagcatg atcaggaagg ccgggtgatt gtggctgaat ttgactcgtt tgtgctggta 2821 acagcatatg tacctaatgc aggccgaggt ctggtacgac tggagtaccg gcagcgctgg 2881 gatgaagcct ttcgcaagtt cctgaagggc ctggcttccc gaaagcccct tgtgctgtgt 2941 ggagacctca atgtggcaca tgaagaaatt gaccttcgca accccaaggg gaacaaaaag 3001 aatgctggct tcacgccaca agagcgccaa ggcttcgggg aattactgca ggctgtgcca 3061 ctggctgaca gctttaggca cctctacccc aacacaccct atgcctacac cttttggact 3121 tatatgatga atgctcgatc caagaatgtt ggttggcgcc ttgattactt tttgttgtcc 3181 cactctctgt tacctgcatt gtgtgacagc aagatccgtt ccaaggccct cggcagtgat 3241 cactgtccta tcaccctata cctagcactg ggatccgact acaaagacca tgacggtgat 3301 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3361 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3421 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3481 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3541 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3601 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3661 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3721 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3781 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3841 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3901 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 3961 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4021 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4081 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4141 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4201 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4261 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4321 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4381 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4441 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4501 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4561 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4621 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4681 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4741 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4801 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4861 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4921 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4981 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5041 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5101 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5161 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5221 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5281 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5341 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5401 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5461 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5521 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5581 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5641 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5701 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5761 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5821 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 5881 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 5941 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6001 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6061 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6121 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6181 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6241 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6301 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6361 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6421 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6481 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6541 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6601 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7441 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8761 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8821 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8881 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8941 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9001 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9061 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9121 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9181 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9241 gctg //