LOCUS V044671 8230 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044671 VERSION V044671 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3084 /label="LASP1(NM_006148)" /note="LASP1(NM_006148)" /gene="LASP1" CDS 3091..3156 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3200..3411 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3424..3692 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3719..4318 /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 4325..4913 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4987..5220 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5292..5413 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5432..5567 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5605..5621) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5629..5645 /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(5653..5683) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5698..5719 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6007..6595) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6766..7626) /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(7627..7731) /gene="bla" /label="AmpR promoter" primer_bind 8205..8221 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgaacccc aactgcgccc ggtgcggcaa gatcgtgtat 2341 cccacggaga aggtgaactg tctggataag ttctggcata aagcatgctt ccattgcgag 2401 acctgcaaga tgacactgaa catgaagaac tacaagggct acgagaagaa gccctactgc 2461 aacgcacact accccaagca gtccttcacc atggtggcgg acaccccgga aaaccttcgc 2521 ctcaagcaac agagtgagct ccagagtcag gtgcgctaca aggaggagtt tgagaagaac 2581 aagggcaaag gtttcagcgt agtggcagac acgcccgagc tccagagaat caagaagacc 2641 caggaccaga tcagtaacat aaaataccat gaggagtttg agaagagccg catgggccct 2701 agcgggggcg agggcatgga gccagagcgt cgggattcac aggacggcag cagctaccgg 2761 cggcccctgg agcagcagca gcctcaccac atcccgacca gtgccccggt ttaccagcag 2821 ccccagcagc agccggtggc ccagtcctat ggtggctaca aggagcctgc agccccagtc 2881 tccatacagc gcagcgcccc aggtggtggc gggaagcggt accgcgcggt gtatgactac 2941 agcgccgccg acgaggacga ggtctccttc caggacgggg acaccatcgt caacgtgcag 3001 cagatcgacg acggctggat gtacgggacg gtggagcgca ccggcgacac ggggatgctg 3061 ccggccaact acgtggaggc catcggatcc gactacaaag accatgacgg tgattataaa 3121 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3181 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3241 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3301 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3361 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3421 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3481 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3541 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3601 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3661 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 3721 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 3781 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 3841 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 3901 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 3961 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4021 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4081 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4141 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4201 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4261 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 4321 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4381 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4441 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4501 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 4561 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 4621 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 4681 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 4741 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 4801 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 4861 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 4921 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 4981 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5041 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5101 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5161 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5221 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5281 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5401 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 5461 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 5521 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 5581 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 5641 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 5701 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 5761 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 5821 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 5881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6181 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6421 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 6541 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 6601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 6661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 6721 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 6781 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 6841 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6901 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6961 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7021 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7081 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7141 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7201 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7261 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7321 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7381 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7441 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7501 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 7561 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 7621 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 7681 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 7741 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 7801 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 7861 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 7921 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 7981 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8041 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8101 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8161 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8221 tgccaagctg //