LOCUS V036068 8701 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036068 VERSION V036068 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..2727 /label="Ndufb7(NM_025843)" /note="Ndufb7(NM_025843)" /gene="Ndufb7" CDS 2734..2799 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2843..3054 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3067..3335 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3364..3373 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3394..4056 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 4126..4179 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 4180..4779 /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 4780..5368 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5442..5675 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5747..5868 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5908..6043 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6076..6092) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6100..6116 /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(6124..6154) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6169..6190 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6478..7066) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7237..8097) /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(8098..8202) /gene="bla" /label="AmpR promoter" primer_bind 8676..8692 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg gggcgcacct gacccggcgc 2341 tatctgtggg atgcctcggt ggagcctgac cccgagaaga tacccagctt cccgccggac 2401 ctcggctttc cggagcgcaa ggagcgagtg atggtggcca cacaacaaga gatgatggat 2461 gcccagctga cactgcagca acgtgactac tgtgcccact acctcatccg gctgctgaag 2521 tgcaagcgag acagcttccc caacttcctg gcctgcaagc acgagcagca cgactgggac 2581 tactgcgagc acctggatta cgtgaaacgc atgaaggagt ttgaacgtga gcggcgactg 2641 ctccaaagga agaagcgaag ggcactgaag gaagcacggg tagcccaagg ccagggagaa 2701 ggagaggtgg gccctgaggt ggccctggga tccgactaca aagaccatga cggtgattat 2761 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc 2821 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 2881 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 2941 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3001 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3061 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 3121 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 3181 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 3241 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 3301 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 3361 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 3421 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 3481 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 3541 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 3601 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 3661 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 3721 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 3781 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 3841 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 3901 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 3961 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 4021 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 4081 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 4141 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 4201 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4261 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 4321 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 4381 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 4441 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4501 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 4561 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 4621 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 4681 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 4741 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 4801 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4861 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4921 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 4981 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5041 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5101 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5161 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt 5221 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5281 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 5341 cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc 5401 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc 5461 ccaacgaaga taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg 5521 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 5581 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 5641 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 5701 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc 5761 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 5821 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct 5881 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 5941 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 6001 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac 6061 caaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 6121 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 6181 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 6241 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 6301 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 6361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 6421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 6481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 6541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 6601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 6661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 6721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6901 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 7021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 7081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 7141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 7201 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 7261 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 7321 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 7381 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 7441 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 7501 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 7561 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 7621 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 7681 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 7741 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 7801 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 7861 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7921 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7981 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 8041 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 8101 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 8161 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 8221 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat 8281 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 8341 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 8401 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca 8461 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 8521 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt 8581 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag 8641 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct 8701 g //