LOCUS V045047 7993 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045047 VERSION V045047 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..2625 /label="CISD1(NM_018464)" /note="CISD1(NM_018464)" /gene="CISD1" CDS 2632..2658 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2662..2688 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2692..2718 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 2762..2973 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2986..3254 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3287..4081 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 4088..4676 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4750..4983 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5055..5176 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5195..5330 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5368..5384) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5392..5408 /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(5416..5446) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5461..5482 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5770..6358) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6529..7389) /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(7390..7494) /gene="bla" /label="AmpR promoter" primer_bind 7968..7984 /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 catgagtctg acttccagtt ccagcgtacg agttgaatgg 2341 atcgcagcag ttaccattgc tgctgggaca gctgcaattg gttatctagc ttacaaaaga 2401 ttttatgtta aagatcatcg aaataaagct atgataaacc ttcacatcca gaaagacaac 2461 cccaagatag tacatgcttt tgacatggag gatttgggag ataaagctgt gtactgccgt 2521 tgttggaggt ccaaaaagtt cccattctgt gatggggctc acacaaaaca taacgaagag 2581 actggagaca atgtgggccc tctgatcatc aagaaaaaag aaactggatc ctacccatac 2641 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat 2701 gatgtcccag actacgcata gcggccgcga aggatctgcg atcgctccgg tgcccgtcag 2761 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga 2821 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc 2881 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt 2941 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct 3001 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg 3061 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc 3121 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc 3181 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg 3241 cgccgttaca gatccaagct gtgaccggcg cctacgctag gccaccatgg ctgaacaaga 3301 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 3361 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 3421 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc 3481 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 3541 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 3601 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 3661 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 3721 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 3781 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt 3841 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 3901 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 3961 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 4021 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 4081 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 4141 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 4201 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 4261 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 4321 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 4381 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 4441 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 4501 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 4561 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 4621 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 4681 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 4741 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 4801 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 4861 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 4921 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 4981 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 5041 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 5101 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 5161 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 5221 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 5281 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 5341 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 5401 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 5461 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 5521 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 5581 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 5641 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5701 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5761 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5821 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 5881 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5941 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6001 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6061 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6181 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 6241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6601 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6721 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6961 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7021 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7141 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7201 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7261 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7321 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7381 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7441 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7501 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 7561 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 7621 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 7681 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 7741 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 7801 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 7861 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 7921 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 7981 cagtgccaag ctg //