LOCUS V022883 9482 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022883 VERSION V022883 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..3525 /label="Trim59(NM_025863)" /note="Trim59(NM_025863)" /gene="Trim59" CDS 3532..3597 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3641..3852 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3865..4133 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4162..4171 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4192..4854 /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 4924..4977 /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 4978..5577 /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 5578..6165 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6239..6472 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6544..6665 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6684..6819 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6857..6873) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6881..6897 /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(6905..6935) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6950..6971 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7259..7847) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8018..8878) /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(8879..8983) /gene="bla" /label="AmpR promoter" primer_bind 9457..9473 /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 gccaccatgc acaattttga ggaggagtta 2341 acgtgtccca tctgttacag catctttgaa gatccccgtg tactaccatg ctctcataca 2401 ttttgtagaa actgtttgga aaatgttctt caggcatctg gtaactttta catatggagg 2461 cctctgcgaa ttccactcaa gtgtcctaac tgcagaagta ttattgaaat tgcttctact 2521 ggcatagaat ccttacctgt aaactttgca ttacgagcaa ttattgaaaa gtaccagcaa 2581 gaagaccacc cagatgttgt cacctgccct gaacactaca ggcaaccatt aaatgtttat 2641 tgtctactag ataagaaatt agtttgtggc cattgtctta ctataggcca acatcatggc 2701 catcctatag atgaccttca aagtgcctat ctgaaagaaa aggatacacc tcagaagttg 2761 ttgaaacagt taactgacac acattggaca gatatcactc gccttattga aaagcttgaa 2821 gaacagaaat gtcattctga gaaaatagtt caaggtgata aggaagttgt tctccagtat 2881 tttaaggagc taattgatac attagaacag aaaaaaaagt attttttggc agctctttgt 2941 gatgttggca aaatgattaa tcaagaatac actccacaga ttcaaggaat gaaggagata 3001 agggaacaac aacttgaatt aatgacaata acgacatctt tacaagatga gtctccactt 3061 aaatttctgg aaaaaattga tgaagtccgc caacgtgtgc agatgttgaa acagagacca 3121 cttcctgagg tccagcctgt tgaaatttat cctcgagtaa gcaatgtatt aaaagaagag 3181 tggagcagaa tagaaattgg acgtattaag aaagctgtta ttcctgaaat gagagtttct 3241 tcaaaaagaa cgccatgttc ctggtctgat aatgatgaaa aagaaatgga actttttaaa 3301 attttaaaca ttgctatagt ttcactcatt tcagtgatac taatgttgat actccttttc 3361 aaccatcaca taataacatt cttaaatgaa atcacttcaa tatgtttctc tgaagtcttt 3421 ctctctgttt accagagttt atctaagaac ttatatgact taaacaatac agtgtgttac 3481 actttatatt tgttaaagga atttatgtgg aaaatagttt ctcgtggatc cgactacaaa 3541 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3601 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3661 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 3721 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 3781 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 3841 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3901 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 3961 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 4021 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4081 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 4141 tgaccggcgc ctacgctaga cgccaccatg gagagcgacg agagcggcct gcccgccatg 4201 gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct ggtgggcggc 4261 ggagagggca cccccaagca gggccgcatg accaacaaga tgaagagcac caaaggcgcc 4321 ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta ccacttcggc 4381 acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg cggctacacc 4441 aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc 4501 tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt ccccgaggac 4561 agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca cctgcacccc 4621 atgggcgata acgtgctggt gggcagcttc gcccgcacct tcagcctgcg cgacggcggc 4681 tactacagct tcgtggtgga cagccacatg cacttcaaga gcgccatcca ccccagcatc 4741 ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggagctgca cagcaacacc 4801 gagctgggca tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc 4861 cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc caccggatct 4921 cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 4981 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 5041 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 5101 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5161 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5221 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5281 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5341 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5401 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5461 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5521 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat 5581 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 5641 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 5701 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 5761 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 5821 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 5881 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 5941 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 6001 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 6061 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc 6121 cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgcctggt acctttaaga 6181 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 6241 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 6301 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 6361 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 6421 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6481 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6541 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6601 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6661 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 6721 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 6781 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 6841 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 6901 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6961 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 7021 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 7081 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 7141 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 7201 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 7261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 7621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 7681 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 7741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 7801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 7861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 7921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 8041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 8101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 8161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 8221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 8281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 8341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 8401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 8461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 8521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 8581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 8641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 8701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 8761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 8821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 8881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 8941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 9001 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 9061 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 9121 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 9181 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 9241 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 9301 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 9361 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 9421 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 9481 tg //