LOCUS V045409 9182 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045409 VERSION V045409 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..3183 /label="PGAM5(NM_001170543)" /note="PGAM5(NM_001170543)" /gene="PGAM5" CDS 3190..3216 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3220..3246 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3250..3276 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3341..3552 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3565..3833 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3862..3871 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3892..4554 /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 4624..4677 /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 4678..5277 /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 5278..5865 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5939..6172 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6244..6365 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6384..6519 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6557..6573) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6581..6597 /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(6605..6635) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6650..6671 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6959..7547) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7718..8578) /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(8579..8683) /gene="bla" /label="AmpR promoter" primer_bind 9157..9173 /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 cgttccggca ggcgctgcag 2341 ctggcggcct gcgggctggc cgggggctcg gccgccgtgc tcttctcggc cgtggcggta 2401 gggaagccgc gcgcaggcgg ggacgcggag ccacgcccgg ctgagccgcc ggcctgggcg 2461 gggggcgcgc ggccgggccc cggtgtctgg gaccccaact gggacaggcg agaaccactg 2521 tctctgatca acgtgcggaa gaggaacgtg gaatctgggg aagaagagct ggcgtccaag 2581 ctggaccact acaaagccaa ggccacgcgg cacatcttcc tcatcaggca ttcccagtac 2641 cacgtggatg gctccctgga gaaggaccgc actctgaccc cgctgggtcg ggagcaggct 2701 gaactcactg ggctccgcct ggcaagcttg gggttgaagt ttaataaaat cgtccattcg 2761 tctatgacgc gcgccataga gaccaccgat atcatcagcc ggcacctgcc aggcgtctgc 2821 aaagtcagca cagatctgct gcgggaaggc gcccccatcg agccagaccc gcccgtgtct 2881 cattggaagc cggaagctgt gcagtattac gaagacggag cccggatcga ggccgccttc 2941 cggaactaca tccaccgcgc agatgccagg caggaggagg acagttacga gatcttcatc 3001 tgtcacgcca acgtcatccg ctacatcgtg tgcagagcac tgcagtttcc tcctgaaggc 3061 tggctccggc tctccctcaa taatggcagc atcacccacc tggtgatccg acccaacggc 3121 cgagttgcgc tcaggaccct cggggacacg gggttcatgc ctcccgacaa gatcactcga 3181 tccctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat 3241 tatgctagct atccttatga tgtcccagac tacgcatagg gatccctgcg atatccagcg 3301 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3361 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 3421 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 3481 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 3541 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3601 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 3661 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 3721 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 3781 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 3841 tgaccggcgc ctacgctaga cgccaccatg gagagcgacg agagcggcct gcccgccatg 3901 gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct ggtgggcggc 3961 ggagagggca cccccaagca gggccgcatg accaacaaga tgaagagcac caaaggcgcc 4021 ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta ccacttcggc 4081 acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg cggctacacc 4141 aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc 4201 tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt ccccgaggac 4261 agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca cctgcacccc 4321 atgggcgata acgtgctggt gggcagcttc gcccgcacct tcagcctgcg cgacggcggc 4381 tactacagct tcgtggtgga cagccacatg cacttcaaga gcgccatcca ccccagcatc 4441 ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggagctgca cagcaacacc 4501 gagctgggca tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc 4561 cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc caccggatct 4621 cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 4681 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 4741 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 4801 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 4861 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 4921 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 4981 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5041 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5101 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5161 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5221 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat 5281 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 5341 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 5401 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 5461 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 5521 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 5581 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 5641 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 5701 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 5761 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc 5821 cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgcctggt acctttaaga 5881 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 5941 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 6001 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 6061 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 6121 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6181 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6241 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6301 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6361 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 6421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 6481 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 6541 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 6601 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 6721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 6781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 6841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 6901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 6961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 7321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 7381 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 7441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 7501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 7561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 7621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7681 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 7741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 7801 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 7861 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 7921 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 7981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 8041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 8101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 8161 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 8221 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 8281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 8341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 8401 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 8461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 8521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 8581 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 8641 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 8701 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 8761 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 8821 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 8881 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 8941 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 9001 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 9061 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 9121 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 9181 tg //