LOCUS V022966 9548 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022966 VERSION V022966 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..3570 /label="Actr3(NM_001205385)" /note="Actr3(NM_001205385)" /gene="Actr3" CDS 3577..3642 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3707..3918 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3931..4199 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4228..4237 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4258..4920 /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 4990..5043 /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 5044..5643 /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 5644..6231 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6305..6538 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6610..6731 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6750..6885 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6923..6939) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6947..6963 /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(6971..7001) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7016..7037 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7325..7913) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8084..8944) /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(8945..9049) /gene="bla" /label="AmpR promoter" primer_bind 9523..9539 /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 cgggacggct gccggcctgt 2341 gtggtggact gtggcacggg atatacaaaa ctaggatatg ctggaaatac agagccacag 2401 tttatcatcc catcatgtat tgccattaaa gagtctgcaa aagtgggtga ccaagcccag 2461 aggagggtga tgaaaggcgt ggatgaccta gacttcttca ttggtgatga agcaatagaa 2521 aagcccacat atgcaacaaa gtggccaatt cgccatggta tagttgaaga ctgggactta 2581 atggaaaggt ttatggagca agtgattttt aaatatttaa gggcagaacc tgaagatcat 2641 tactttcttt tgactgaacc tccactgaat actccagaaa acagggaata tactgctgaa 2701 ataatgtttg aatccttcaa tgttccaggc ttgtacattg ctgtgcaggc tgttcttgcc 2761 ttagctgcat cctggacctc aagacaagta ggagagcgga cgctgacggg tacagtaata 2821 gacagtggag acggagtcac tcatgtcatt cctgtggctg aaggatatgt tatcggcagc 2881 tgtattaaac acattccaat cgcaggaaga gatataacat attttattca gcaactgctg 2941 cgagaccgag aagtaggaat ccctcctgag cagtccttgg aaactgcgaa agcagtgaag 3001 gaacgctaca gttatgtctg cccagattta gtaaaagagt ttaacaagta tgacaccgat 3061 gggtcaaagt ggatcaaaca gtacaccgga gtcaacgcca tctcaaagaa agagttttct 3121 attgatgttg gctatgagcg attcctggga cccgagatct ttttccatcc agagtttgct 3181 aatccagatt ttacacaacc tatctcagaa gttgtagatg aagtcattca gaattgcccc 3241 attgatgtcc ggcgtcctct ctacaagaac attgtcctct ctggtggttc aaccatgttc 3301 agggactttg gacgtcgttt gcaaagagat ttgaagagaa ctgtagatgc caggctgaag 3361 ttaagcgagg agctgagtgg tggtagattg aagcccaagc ctattgatgt acaagttatt 3421 acacaccata tgcagcggta tgcagtctgg tttggagggt caatgctggc ttccacgcct 3481 gagttctacc aagtatgcca caccaaaaag gattatgaag aaattggacc tagcatttgt 3541 cgtcacaatc cagtgtttgg agtcatgtcc ctcgaggact acaaagacca tgacggtgat 3601 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagggatc cctgcgatat 3661 ccagcgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3721 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 3781 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 3841 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 3901 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 3961 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 4021 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4081 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4141 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4201 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4261 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4321 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4381 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4441 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4501 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4561 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4621 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4681 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4741 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 4801 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 4861 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 4921 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 4981 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 5041 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5101 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5161 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5221 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5281 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5341 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5401 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5461 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5521 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5581 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5641 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5701 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5761 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5821 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5881 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5941 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 6001 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 6061 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6121 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6181 ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct 6241 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6301 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6361 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6421 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6481 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6541 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6601 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6721 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 6781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 6841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 6901 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7741 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 7801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8041 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8101 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8161 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8221 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8281 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8341 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8401 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8461 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8521 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8581 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8641 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8701 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8761 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8821 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8881 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8941 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9001 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9061 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9121 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9181 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9241 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9301 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9361 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9421 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9481 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9541 ccaagctg //