LOCUS V032079 7844 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032079 VERSION V032079 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 1960..2171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2184..2452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 2498..2725 /label="Cd24a(NM_009846)" /note="Cd24a(NM_009846)" /gene="Cd24a" CDS 2732..2797 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2813..3312 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 3333..3932 /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 3939..4527 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4601..4834 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4906..5027 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5046..5181 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5219..5235) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5243..5259 /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(5267..5297) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5312..5333 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5621..6209) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6380..7240) /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(7241..7345) /gene="bla" /label="AmpR promoter" primer_bind 7819..7835 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca 1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc 2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta 2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct 2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc 2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc 2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt 2461 gaccggcgcc tactctagag ctagcgaatt cgccaccatg ggcagagcga tggtggccag 2521 gctagggctg gggttgctgc ttctggcact gctcctaccc acgcagattt actgcaacca 2581 aacatctgtt gcaccgtttc ccggtaacca gaatatttct gcttccccaa atccaagtaa 2641 cgctaccacc agagggggtg gcagctccct gcagtccaca gctggtctcc tggctctctc 2701 tctctctctt ctacatctct actgtctcga ggactacaaa gaccatgacg gtgattataa 2761 agatcatgac atcgactaca aggatgacga tgacaagtag tgagcggccg ctgggtaggg 2821 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg 2881 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg 2941 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc 3001 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact 3061 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc 3121 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc 3181 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc 3241 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct 3301 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc 3361 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact 3421 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc 3481 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 3541 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 3601 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 3661 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 3721 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 3781 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 3841 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 3901 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa 3961 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 4021 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 4081 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 4141 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 4201 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 4261 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 4321 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 4381 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 4441 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 4501 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 4561 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 4621 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 4681 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 4741 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 4801 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 4861 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 4921 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 4981 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 5041 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 5101 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 5161 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 5221 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5281 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5341 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5401 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5461 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5521 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5581 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5641 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5701 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5761 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5821 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5881 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5941 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6001 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6061 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6121 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6181 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6241 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6301 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6361 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6421 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6481 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6541 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6601 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6661 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6721 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6781 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6841 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6901 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6961 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7021 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7081 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7141 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7201 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7261 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7321 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 7381 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 7441 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 7501 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 7561 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 7621 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 7681 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 7741 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 7801 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //