LOCUS V020435 9488 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020435 VERSION V020435 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2289..3530 /label="NR2F2(NM_021005)" /note="NR2F2(NM_021005)" /gene="NR2F2" CDS 3537..3602 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3647..3858 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3871..4139 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4168..4177 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4198..4860 /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 4930..4983 /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 4984..5583 /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 5584..6171 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6245..6478 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6550..6671 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6690..6825 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6863..6879) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6887..6903 /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(6911..6941) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6956..6977 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7265..7853) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8024..8884) /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(8885..8989) /gene="bla" /label="AmpR promoter" primer_bind 9463..9479 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagccaccat ggcaatggta gtcagcacgt ggcgcgaccc ccaggacgag gtgcccggct 2341 cacagggcag ccaggcctcg caggcgccgc ccgtgcccgg cccgccgccc ggcgccccgc 2401 acacgccaca gacgcccggc caagggggcc cagccagcac gccagcccag acggcggccg 2461 gtggccaggg cggccctggc ggcccgggta gcgacaagca gcagcagcag caacacatcg 2521 agtgcgtggt gtgcggagac aagtcgagcg gcaagcacta cggccagttc acgtgcgagg 2581 gctgcaagag cttcttcaag cgcagcgtgc ggaggaacct gagctacacg tgccgcgcca 2641 accggaactg tcccatcgac cagcaccatc gcaaccagtg ccagtactgc cgcctcaaaa 2701 agtgcctcaa agtgggcatg agacgggaag cggtgcagag gggcaggatg ccgccgaccc 2761 agccgaccca cgggcagttc gcgctgacca acggggatcc cctcaactgc cactcgtacc 2821 tgtccggata tatttccctg ctgttgcgcg cggagcccta tcccacgtcg cgcttcggca 2881 gccaatgcat gcagcccaac aacatcatgg gtatcgagaa catttgcgaa ctggccgcga 2941 ggatgctctt cagcgccgtc gagtgggccc ggaacatccc cttcttcccc gacctgcaga 3001 tcacggacca ggtggccctg cttcgcctca cctggagcga gctgtttgtg ttgaatgcgg 3061 cgcagtgctc catgcccctc cacgtcgccc cgctcctggc cgccgccggc ctgcatgctt 3121 cgcccatgtc cgccgaccgg gtggtcgcct ttatggacca catacggatc ttccaagagc 3181 aagtggagaa gctcaaggcg ctgcacgttg actcagccga gtacagctgc ctcaaggcca 3241 tagtcctgtt cacctcagat gcctgtggtc tctctgatgt agcccatgtg gaaagcttgc 3301 aggaaaagtc tcagtgtgct ttggaagaat acgttaggag ccagtacccc aaccagccga 3361 cgagattcgg aaagcttttg cttcgcctcc cttccctccg caccgtctcc tcctcagtca 3421 tagagcaatt gtttttcgtc cgtttggtag gtaaaacccc catcgaaacc ctcatccggg 3481 atatgttact gtccggcagc agttttaact ggccgtatat ggcaattcaa ggatccgact 3541 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca 3601 agtgagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3661 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 3721 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 3781 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 3841 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 3901 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 3961 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4021 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4081 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4141 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4201 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4261 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4321 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4381 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4441 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4501 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4561 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4621 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4681 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 4741 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 4801 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 4861 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 4921 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 4981 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5041 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5101 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5161 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5221 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5281 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5341 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5401 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5461 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5521 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5581 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5641 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5701 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5761 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5821 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5881 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 5941 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 6001 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6061 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6121 ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct 6181 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6241 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6301 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6361 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6421 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6481 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6541 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6661 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 6721 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 6781 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 6841 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6901 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 6961 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7021 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7081 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7141 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7201 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7261 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7321 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7381 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7441 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7501 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7561 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7621 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7681 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 7741 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7801 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7861 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7921 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7981 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8041 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8101 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8161 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8221 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8281 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8341 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8401 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8461 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8521 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8581 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8641 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8701 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8761 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8821 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8881 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8941 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9001 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9061 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9121 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9181 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9241 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9301 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9361 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9421 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9481 ccaagctg //