LOCUS V043125 9136 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043125 VERSION V043125 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..3162 /label="SNRPA(NM_004596)" /note="SNRPA(NM_004596)" /gene="SNRPA" CDS 3169..3234 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3278..3489 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3502..3770 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3799..3808 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3829..4491 /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 4561..4614 /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 4615..5214 /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 5215..5803 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5877..6110 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6182..6303 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6343..6478 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6511..6527) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6535..6551 /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(6559..6589) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6604..6625 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6913..7501) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7672..8532) /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(8533..8637) /gene="bla" /label="AmpR promoter" primer_bind 9111..9127 /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 cagttcccga gacccgccct 2341 aaccacacta tttatatcaa caacctcaat gagaagatca agaaggatga gctaaaaaag 2401 tccctgtacg ccatcttctc ccagtttggc cagatcctgg atatcctggt atcacggagc 2461 ctgaagatga ggggccaggc ctttgtcatc ttcaaggagg tcagcagcgc caccaacgcc 2521 ctgcgctcca tgcagggttt ccctttctat gacaaaccta tgcgtatcca gtatgccaag 2581 accgactcag atatcattgc caagatgaaa ggcaccttcg tggagcggga ccgcaagcgg 2641 gagaagagga agcccaagag ccaggagacc ccggccacca agaaggctgt gcaaggcggg 2701 ggagccaccc ccgtggtggg ggctgtccag gggcctgtcc cgggcatgcc gccgatgact 2761 caggcgcccc gcattatgca ccacatgccg ggccagccgc cctacatgcc gccccctggt 2821 atgatccccc cgccaggcct tgcacctggc cagatcccac caggggccat gcccccgcag 2881 cagcttatgc caggacagat gccccctgcc cagcctcttt ctgagaatcc accgaatcac 2941 atcttgttcc tcaccaacct gccagaggag accaacgagc tcatgctgtc catgcttttc 3001 aatcagttcc ctggcttcaa ggaggtccgt ctggtacccg ggcggcatga catcgccttc 3061 gtggagtttg acaatgaggt acaggcaggg gcagctcgcg atgccctgca gggctttaag 3121 atcacgcaga acaacgccat gaagatctcc tttgccaaga agggatccga ctacaaagac 3181 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3241 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 3301 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 3361 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 3421 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 3481 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 3541 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 3601 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 3661 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 3721 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 3781 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 3841 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 3901 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 3961 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4021 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4081 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 4141 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4201 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4261 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 4321 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 4381 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 4441 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 4501 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 4561 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 4621 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4681 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4741 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 4801 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 4861 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 4921 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 4981 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5041 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5101 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5161 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5221 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5281 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5341 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5401 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5461 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5521 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5581 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 5641 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 5701 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 5761 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 5821 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 5881 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 5941 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6001 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6061 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6121 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6181 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6241 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6301 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 6361 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6421 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6481 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 6541 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6601 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6661 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6721 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7321 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7621 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7981 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8281 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8521 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8641 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8701 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8761 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8821 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8881 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 8941 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9001 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9061 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9121 ggccagtgcc aagctg //