LOCUS V042387 8971 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042387 VERSION V042387 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 2303..3061 /label="C1qb(NM_009777)" /note="C1qb(NM_009777)" /gene="C1qb" CDS 3062..3079 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 3130..3341 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3354..3622 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3651..3660 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3681..4343 /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 4413..4466 /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 4467..5066 /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 5067..5654 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5728..5961 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6033..6154 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6173..6308 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6346..6362) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6370..6386 /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(6394..6424) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6439..6460 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6748..7336) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7507..8367) /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(8368..8472) /gene="bla" /label="AmpR promoter" primer_bind 8946..8962 /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 gaattcgcca ccatgaagac acagtggggt gaggtctgga cacacctgtt 2341 actgctgctt ctaggttttc tccatgtgtc ctgggcccaa agcagctgca ccgggccccc 2401 tggcatccct ggcatccctg gggtccctgg ggttcctggc tctgatggcc aaccaggcac 2461 tccagggata aagggggaga aagggctccc tggactggct ggagaccttg gtgagtttgg 2521 agagaaaggg gacccaggga tccctgggac tccaggcaaa gttggcccta agggtcccgt 2581 cggccctaag ggtactccag gcccctctgg accccgcggt cccaaaggcg attctgggga 2641 ctacggggct acacagaaag tcgccttctc tgccctgagg accatcaaca gccccttgcg 2701 accgaaccag gtcattcgct tcgaaaaggt gatcaccaac gcgaacgaga actatgagcc 2761 acgcaacggc aagttcacct gcaaggtgcc tggcctctac tacttcacct atcatgccag 2821 ctcccggggc aacctgtgtg tgaatctcgt tcgtggccgc gatcgggaca gcatgcagaa 2881 agtagtcacc ttctgtgact atgcccagaa caccttccag gtgaccacag gtggggtagt 2941 cttgaagcta gagcaagagg aggttgttca cctgcaggcc acagacaaga actccctcct 3001 gggcattgag ggtgccaaca gcatcttcac tggctttctg cttttccctg acatggatgc 3061 gcatcaccat catcaccatt gaggatccgc ggccgcgaag gatctgcgat cgctccggtg 3121 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 3181 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 3241 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 3301 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc 3361 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc 3421 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt 3481 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca 3541 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt 3601 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg 3661 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga 3721 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga 3781 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg 3841 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc 3901 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg 3961 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca 4021 aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca 4081 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg 4141 cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc 4201 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc 4261 gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct 4321 tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg 4381 gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg 4441 gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca 4501 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 4561 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 4621 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 4681 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 4741 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 4801 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 4861 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 4921 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 4981 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 5041 gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat 5101 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 5161 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 5221 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 5281 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 5341 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 5401 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 5461 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt 5521 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 5581 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctcccttt 5641 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 5701 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 5761 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 5821 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5881 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 5941 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6001 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 6061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6121 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 6181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6241 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6301 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6361 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6421 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6481 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8461 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8521 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8581 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8641 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8701 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8761 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8821 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8881 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8941 acgttgtaaa acgacggcca gtgccaagct g //