LOCUS V039697 8905 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039697 VERSION V039697 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..2931 /label="Rab1a(NM_008996)" /note="Rab1a(NM_008996)" /gene="Rab1a" CDS 2938..3003 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3047..3258 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3271..3539 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3568..3577 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3598..4260 /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 4330..4383 /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 4384..4983 /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 4984..5572 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5646..5879 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5951..6072 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6112..6247 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6280..6296) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6304..6320 /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(6328..6358) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6373..6394 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6682..7270) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7441..8301) /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(8302..8406) /gene="bla" /label="AmpR promoter" primer_bind 8880..8896 /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 gccaccatgt ccagcatgaa tcccgaatat 2341 gattatttat tcaagttact tctgattggc gattctgggg ttggaaagtc ctgccttctc 2401 cttaggtttg cagatgatac gtatacggaa agctacatca gcacaattgg tgtggatttc 2461 aagatacgaa ctatagagtt agatgggaaa acaatcaagc tacagatatg ggacacagca 2521 ggccaggaaa gatttcgaac aatcacttcc agttattaca gaggagccca tggcatcata 2581 gttgtgtatg atgtgacaga tcaggagtcc ttcaataacg ttaaacagtg gctgcaggag 2641 atagatcgct acgccagtga aaatgtcaac aagttgttgg tagggaacaa atgtgacctg 2701 accacaaaga aagtagtaga ctacacaaca gcaaaggaat ttgcagattc ccttggaatt 2761 ccatttttgg aaaccagtgc taagaacgca acgaatgtag aacagtcttt catgacgatg 2821 gcagctgaga ttaaaaagcg aatgggtcct ggagctacag ctggtggtgc cgagaagtcc 2881 aatgttaaaa tccagagcac tccagtcaag cagtcaggtg gaggctgctg cggatccgac 2941 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 3001 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3061 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 3121 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 3181 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 3241 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 3301 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 3361 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 3421 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 3481 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 3541 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 3601 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 3661 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 3721 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 3781 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 3841 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 3901 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 3961 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4021 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4081 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 4141 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 4201 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 4261 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 4321 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 4381 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 4441 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 4501 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 4561 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 4621 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 4681 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 4741 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 4801 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 4861 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 4921 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 4981 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5041 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5101 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5161 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5221 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5281 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 5341 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 5401 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 5461 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 5521 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 5581 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 5641 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg 5701 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 5761 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 5821 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 5881 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 5941 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6001 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6061 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc 6121 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 6181 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 6241 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg 6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7081 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8401 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 8461 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 8521 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 8581 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 8641 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 8701 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 8761 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 8821 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 8881 taaaacgacg gccagtgcca agctg //