LOCUS V023289 7825 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023289 VERSION V023289 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..2874 /label="ANKRD22(NM_144590)" /note="ANKRD22(NM_144590)" /gene="ANKRD22" CDS 2881..2946 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2990..3201 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3214..3482 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3515..3913 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 3920..4508 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4582..4815 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4887..5008 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5027..5162 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5200..5216) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5224..5240 /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(5248..5278) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5293..5314 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5602..6190) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6361..7221) /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(7222..7326) /gene="bla" /label="AmpR promoter" primer_bind 7800..7816 /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 catgggaatc ctatactctg agcccatctg ccaagcagcc 2341 tatcagaatg actttggaca agtgtggcgg tgggtgaaag aagacagcag ctatgccaac 2401 gttcaagatg gctttaatgg agacacgccc ctgatctgtg cttgcaggcg agggcatgtg 2461 agaatcgttt ccttcctttt aagaagaaat gctaatgtca acctcaaaaa ccagaaagag 2521 agaacctgct tgcattatgc tgtgaagaaa aaatttacct tcattgatta tctactaatt 2581 atcctcttaa tgcctgttct gcttattggg tatttcctca tggtatcaaa gacaaagcag 2641 aatgaggctc ttgtacgaat gctacttgat gctggcgtcg aagttaatgc tacagattgt 2701 tatggctgta ccgcattaca ttatgcctgt gaaatgaaaa accagtctct tatccctctg 2761 ctcttggaag cccgtgcaga ccccacaata aagaataagc atggtgagag ctcactggat 2821 attgcacgga gattaaaatt ttcccagatt gaattaatgc taaggaaagc attgggatcc 2881 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 2941 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc 3001 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag 3061 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc 3121 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac 3181 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc 3241 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc 3301 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg 3361 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc 3421 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga 3481 tccaagctgt gaccggcgcc tacgctaggc caccatggcc aagcctttgt ctcaagaaga 3541 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta 3601 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata 3661 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc 3721 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc 3781 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa 3841 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt 3901 gtgggagggc taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 3961 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 4021 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 4081 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 4141 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 4201 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 4261 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 4321 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 4381 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 4441 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 4501 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 4561 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc 4621 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 4681 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 4741 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 4801 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 4861 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 4921 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4981 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 5041 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 5101 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 5161 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg 5221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 5281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 5701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5761 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 5821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 5881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 6001 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 6061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 6121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7321 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 7381 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 7441 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 7501 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 7561 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 7621 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 7681 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 7741 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 7801 taaaacgacg gccagtgcca agctg //