LOCUS V033695 7990 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033695 VERSION V033695 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..2844 /label="ARF1(NM_001658)" /note="ARF1(NM_001658)" /gene="ARF1" CDS 2851..2916 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2960..3171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3184..3452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3479..4078 /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 4085..4673 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4747..4980 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5052..5173 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5192..5327 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5365..5381) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5389..5405 /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(5413..5443) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5458..5479 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5767..6355) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6526..7386) /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(7387..7491) /gene="bla" /label="AmpR promoter" primer_bind 7965..7981 /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 catggggaac atcttcgcca acctcttcaa gggccttttt 2341 ggcaaaaaag aaatgcgcat cctcatggtg ggcctggatg ctgcagggaa gaccacgatc 2401 ctctacaagc ttaagctggg tgagatcgtg accaccattc ccaccatagg cttcaacgtg 2461 gaaaccgtgg agtacaagaa catcagcttc actgtgtggg acgtgggtgg ccaggacaag 2521 atccggcccc tgtggcgcca ctacttccag aacacacaag gcctgatctt cgtggtggac 2581 agcaatgaca gagagcgtgt gaacgaggcc cgtgaggagc tcatgaggat gctggccgag 2641 gacgagctcc gggatgctgt cctcctggtg ttcgccaaca agcaggacct ccccaacgcc 2701 atgaatgcgg ccgagatcac agacaagctg gggctgcact cactacgcca caggaactgg 2761 tacattcagg ccacctgcgc caccagcggc gacgggctct atgaaggact ggactggctg 2821 tccaatcagc tccggaacca gaagggatcc gactacaaag accatgacgg tgattataaa 2881 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 2941 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3001 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3061 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3121 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3181 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3241 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3301 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3361 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3421 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 3481 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 3541 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 3601 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 3661 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 3721 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 3781 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 3841 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 3901 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 3961 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4021 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 4081 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4141 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4201 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4261 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 4321 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 4381 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 4441 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 4501 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 4561 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 4621 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 4681 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 4741 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 4801 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 4861 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 4921 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 4981 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5041 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5161 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 5221 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 5281 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 5341 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 5401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 5461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 5521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 5941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6181 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 6301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 6361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 6421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 6481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 6541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 6601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 6781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 6841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 6901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 6961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 7321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 7381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 7441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 7501 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 7561 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 7621 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 7681 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 7741 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 7801 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 7861 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 7921 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 7981 tgccaagctg //