LOCUS V044572 9005 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044572 VERSION V044572 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..3006 /label="LYPLA1(NM_006330)" /note="LYPLA1(NM_006330)" /gene="LYPLA1" CDS 3013..3039 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3043..3069 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3073..3099 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3164..3375 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3388..3656 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3685..3694 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3715..4377 /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 4447..4500 /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 4501..5100 /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 5101..5688 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5762..5995 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6067..6188 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6207..6342 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6380..6396) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6404..6420 /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(6428..6458) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6473..6494 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6782..7370) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7541..8401) /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(8402..8506) /gene="bla" /label="AmpR promoter" primer_bind 8980..8996 /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 gccaccatgt gcggcaataa catgtcaacc 2341 ccgctgcccg ccatcgtgcc cgccgcccgg aaggccaccg ctgcggtgat tttcctgcat 2401 ggattgggag atactgggca cggatgggca gaagcctttg caggtatcag aagttcacat 2461 atcaaatata tctgcccgca tgcgcctgtt aggcctgtta cattaaatat gaacgtggct 2521 atgccttcat ggtttgatat tattgggctt tcaccagatt cacaggagga tgaatctggg 2581 attaaacagg cagcagaaaa tataaaagct ttgattgatc aagaagtgaa gaatggcatt 2641 ccttctaaca gaattatttt gggagggttt tctcagggag gagctttatc tttatatact 2701 gcccttacca cacagcagaa actggcaggt gtcactgcac tcagttgctg gcttccactt 2761 cgggcttcct ttccacaggg tcctatcggt ggtgctaata gagatatttc tattctccag 2821 tgccacgggg attgtgaccc tttggttccc ctgatgtttg gttctcttac ggtggaaaaa 2881 ctaaaaacat tggtgaatcc agccaatgtg acctttaaaa cctatgaagg tatgatgcac 2941 agttcgtgtc aacaggaaat gatggatgtc aagcaattca ttgataaact cctacctcca 3001 attgatctcg agtacccata cgacgtacct gattatgcaa cttatcctta tgatgtacct 3061 gattatgcta gctatcctta tgatgtccca gactacgcat agggatccct gcgatatcca 3121 gcgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3181 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3241 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3301 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3361 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 3421 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 3481 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 3541 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 3601 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 3661 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 3721 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 3781 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 3841 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 3901 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 3961 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 4021 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4081 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4141 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4201 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4261 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4321 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4381 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 4441 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 4501 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 4561 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 4621 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 4681 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 4741 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 4801 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 4861 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 4921 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 4981 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 5041 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5101 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5161 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5221 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5281 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5341 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5401 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5461 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 5521 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 5581 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 5641 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 5701 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5761 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 5821 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5881 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 5941 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6001 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6061 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6121 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6181 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6241 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6301 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6361 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6421 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6481 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6541 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6601 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6661 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6721 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6781 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6961 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7201 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7381 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7441 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 7981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8521 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8581 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 8641 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 8701 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 8761 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8821 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 8881 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 8941 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9001 agctg //