LOCUS V021787 9787 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021787 VERSION V021787 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..3814 /label="FGFRL1(NM_001004356)" /note="FGFRL1(NM_001004356)" /gene="FGFRL1" CDS 3830..3895 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3946..4157 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4170..4438 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4467..4476 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4497..5159 /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 5229..5282 /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 5283..5882 /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 5883..6470 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6544..6777 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6849..6970 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6989..7124 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7162..7178) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7186..7202 /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(7210..7240) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7255..7276 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7564..8152) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8323..9183) /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(9184..9288) /gene="bla" /label="AmpR promoter" primer_bind 9762..9778 /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 ccatgacgcc gagccccctg ttgctgctcc tgctgccgcc 2341 gctgctgctg ggggccttcc cgccggccgc cgccgcccga ggccccccaa agatggcgga 2401 caaggtggtc ccacggcagg tggcccggct gggccgcact gtgcggctgc agtgcccagt 2461 ggagggggac ccgccgccgc tgaccatgtg gaccaaggat ggccgcacca tccacagcgg 2521 ctggagccgc ttccgcgtgc tgccgcaggg gctgaaggtg aagcaggtgg agcgggagga 2581 tgccggcgtg tacgtgtgca aggccaccaa cggcttcggc agcctgagcg tcaactacac 2641 cctcgtcgtg ctggatgaca ttagcccagg gaaggagagc ctggggcccg acagctcctc 2701 tgggggtcaa gaggaccccg ccagccagca gtgggcacga ccgcgcttca cacagccctc 2761 caagatgagg cgccgggtga tcgcacggcc cgtgggtagc tccgtgcggc tcaagtgcgt 2821 ggccagcggg caccctcggc ccgacatcac gtggatgaag gacgaccagg ccttgacgcg 2881 cccagaggcc gctgagccca ggaagaagaa gtggacactg agcctgaaga acctgcggcc 2941 ggaggacagc ggcaaataca cctgccgcgt gtcgaaccgc gcgggcgcca tcaacgccac 3001 ctacaaggtg gatgtgatcc agcggacccg ttccaagccc gtgctcacag gcacgcaccc 3061 cgtgaacacg acggtggact tcggggggac cacgtccttc cagtgcaagg tgcgcagcga 3121 cgtgaagccg gtgatccagt ggctgaagcg cgtggagtac ggcgccgagg gccgccacaa 3181 ctccaccatc gatgtgggcg gccagaagtt tgtggtgctg cccacgggtg acgtgtggtc 3241 gcggcccgac ggctcctacc tcaataagct gctcatcacc cgtgcccgcc aggacgatgc 3301 gggcatgtac atctgccttg gcgccaacac catgggctac agcttccgca gcgccttcct 3361 caccgtgctg ccagacccaa aaccgccagg gccacctgtg gcctcctcgt cctcggccac 3421 tagcctgccg tggcccgtgg tcatcggcat cccagccggc gctgtcttca tcctgggcac 3481 cctgctcctg tggctttgcc aggcccagaa gaagccgtgc acccccgcgc ctgcccctcc 3541 cctgcctggg caccgcccgc cggggacggc ccgcgaccgc agcggagaca aggaccttcc 3601 ctcgttggcc gccctcagcg ctggccctgg tgtggggctg tgtgaggagc atgggtctcc 3661 ggcagccccc cagcacttac tgggcccagg cccagttgct ggccctaagt tgtaccccaa 3721 actctacaca gacatccaca cacacacaca cacacactct cacacacact cacacgtgga 3781 gggcaaggtc caccagcaca tccactatca gtgcggcggt gggggctcag actacaaaga 3841 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtaggg 3901 atccgcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat 3961 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa 4021 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 4081 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 4141 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc 4201 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg 4261 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc 4321 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga 4381 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca 4441 agctgtgacc ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg 4501 ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg 4561 gcggcggaga gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag 4621 gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact 4681 tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct 4741 acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct 4801 accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg 4861 aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc 4921 accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg 4981 gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca 5041 gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca 5101 acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca 5161 gatcccgcgc tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg 5221 gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc 5281 ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg 5341 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 5401 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 5461 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 5521 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 5581 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 5641 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 5701 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 5761 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 5821 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 5881 gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5941 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 6001 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 6061 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 6121 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 6181 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 6241 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 6301 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 6361 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 6421 tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt 6481 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 6541 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 6601 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6661 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6721 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6781 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6841 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6901 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6961 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 7021 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 7081 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 7141 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 7201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7981 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 8041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 8101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 9001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 9061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9301 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9361 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9541 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9601 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9661 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9721 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9781 caagctg //