LOCUS V030014 8522 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030014 VERSION V030014 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..3354 /label="MCU(NM_138357)" /note="MCU(NM_138357)" /gene="MCU" CDS 3361..3387 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3391..3417 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3421..3447 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3492..3703 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3716..3984 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4011..4610 /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 4617..5205 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5279..5512 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5584..5705 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5724..5859 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5897..5913) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5921..5937 /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(5945..5975) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5990..6011 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6299..6887) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7058..7918) /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(7919..8023) /gene="bla" /label="AmpR promoter" primer_bind 8497..8513 /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 catggcggcc gccgcaggta gatcgctcct gctgctcctc 2341 tcctctcggg gcggcggcgg cgggggcgcc ggcggctgcg gggcgctgac tgccggctgc 2401 ttccctgggc tgggcgtcag ccgccaccgg cagcagcagc accaccggac ggtacaccag 2461 aggatcgctt cctggcagaa tttgggagct gtttattgca gcactgttgt gccctctgat 2521 gatgttacag tggtttatca aaatgggtta cctgtgatat ctgtgaggct accatcccgg 2581 cgtgaacgct gtcagttcac actcaagcct atctctgact ctgttggtgt atttttacga 2641 caactgcaag aagaggatcg gggaattgac agagttgcta tctattcacc agatggtgtt 2701 cgcgttgctg cttcaacagg aatagacctc ctcctccttg atgactttaa gctggtcatt 2761 aatgacttaa cataccacgt acgaccacca aaaagagacc tcttaagtca tgaaaatgca 2821 gcaacgctga atgatgtaaa gacattggtc cagcaactat acaccacact gtgcattgag 2881 cagcaccagt taaacaagga aagggagctt attgaaagac tagaggatct caaagagcag 2941 ctggctcccc tggaaaaggt acgaattgag attagcagaa aagctgagaa gaggaccact 3001 ttggtgctat ggggtggcct tgcctacatg gccacacagt ttggcatttt ggcccggctt 3061 acctggtggg aatattcctg ggacatcatg gagccagtaa catacttcat cacttatgga 3121 agtgccatgg caatgtatgc atattttgta atgacacgcc aggaatatgt ttatccagaa 3181 gccagagaca gacaatactt actatttttc cataaaggag ccaaaaagtc acgttttgac 3241 ctagagaaat acaatcaact caaggatgca attgctcagg cagaaatgga ccttaagaga 3301 ctgagagacc cattacaagt acatctgcct ctccgacaaa ttggtgaaaa agatggatcc 3361 tacccatacg acgtacctga ttatgcaact tatccttatg atgtacctga ttatgctagc 3421 tatccttatg atgtcccaga ctacgcatag gcggccgcga aggatctgcg atcgctccgg 3481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 3541 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 3601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 3661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc 3721 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag 3781 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag 3841 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact 3901 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt 3961 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt 4021 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg 4081 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg 4141 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 4201 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 4261 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 4321 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 4381 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 4441 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 4501 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 4561 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc 4621 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 4681 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 4741 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 4801 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 4861 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 4921 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 4981 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct 5041 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 5101 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 5161 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga 5221 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 5281 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 5341 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 5401 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 5461 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 5521 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 5581 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 5641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5701 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 5761 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 5821 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 5881 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 5941 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6001 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 6061 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 6121 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 6181 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 6241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 6301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 6361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 6421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 6481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 6541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 6601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 6661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 6721 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 6781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 6841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 6901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 6961 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 7081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 7141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 7201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 7261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 7321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 7381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 7441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 7501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 7561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 7621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 7681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 7741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 7801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 7861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 7921 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 7981 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 8041 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 8101 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 8161 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 8221 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 8281 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 8341 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 8401 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 8461 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 8521 tg //