LOCUS V042388 7925 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042388 VERSION V042388 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" regulatory 2296..2305 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2305..2370 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2374..2757 /label="UBA52(NM_001321019)" /note="UBA52(NM_001321019)" /gene="UBA52" CDS 2764..2790 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2794..2820 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2824..2850 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 2895..3106 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3119..3387 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3414..4013 /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 4020..4608 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4682..4915 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4987..5108 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5127..5262 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5300..5316) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5324..5340 /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(5348..5378) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5393..5414 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5702..6290) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6461..7321) /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(7322..7426) /gene="bla" /label="AmpR promoter" primer_bind 7900..7916 /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 catggactac aaagaccatg acggtgatta taaagatcat 2341 gacatcgact acaaggatga cgatgacaag cttatgcaga tctttgtgaa gaccctcact 2401 ggcaaaacca tcacccttga ggtcgagccc agtgacacca ttgagaatgt caaagccaaa 2461 attcaagaca aggagggtat cccacctgac cagcagcgtc tgatatttgc cggcaaacag 2521 ctggaggatg gccgcactct ctcagactac aacatccaga aagagtccac cctgcacctg 2581 gtgttgcgcc tgcgaggtgg cattattgag ccttctctcc gccagcttgc ccagaaatac 2641 aactgcgaca agatgatctg ccgcaagtgc tatgctcgcc ttcaccctcg tgctgtcaac 2701 tgccgcaaga agaagtgtgg tcacaccaac aacctgcgtc ccaagaagaa ggtcaaagga 2761 tcctacccat acgacgtacc tgattatgca acttatcctt atgatgtacc tgattatgct 2821 agctatcctt atgatgtccc agactacgca taggcggccg cgaaggatct gcgatcgctc 2881 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 2941 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 3001 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 3061 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 3121 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 3181 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 3241 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 3301 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 3361 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 3421 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 3481 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 3541 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 3601 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 3661 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 3721 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 3781 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 3841 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 3901 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 3961 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 4021 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 4081 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 4141 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 4201 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 4261 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 4321 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 4381 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 4441 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 4501 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 4561 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 4621 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 4681 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 4741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 4801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 4861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 4921 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 4981 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 5041 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 5101 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 5161 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 5221 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 5281 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 5341 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 5401 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 5461 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 5521 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5581 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 6001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 6061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 6121 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 6181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 6241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 6301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 7021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 7081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 7141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 7201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 7261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 7321 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7381 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7441 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 7501 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 7561 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 7621 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 7681 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 7741 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 7801 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 7861 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 7921 agctg //