LOCUS V037543 8709 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037543 VERSION V037543 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..2617 /label="Rpl36(NM_018730)" /note="Rpl36(NM_018730)" /gene="Rpl36" promoter 2662..2873 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2886..3154 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3183..3192 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3213..3875 /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 3945..3998 /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 4005..4799 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 4806..5394 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5466..5699 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5771..5892 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5911..6046 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6084..6100) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6108..6124 /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(6132..6162) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6177..6198 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6486..7074) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7245..8105) /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(8106..8210) /gene="bla" /label="AmpR promoter" primer_bind 8684..8700 /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 ccatggctct gcgctacccc atggccgtgg gcctcaacaa 2341 gggccacaag gtgacgaaaa acgtcagtaa gccgagacac agccggcgac gcgggcgcct 2401 caccaaacac accaagttcg tgcgggacat gatccgggag gtgtgcggct tcgcacccta 2461 cgagcggcga gccatggagt tgctcaaagt gtccaaggac aagcgcgcac tcaagttcat 2521 caagaagagg gtgggcacgc acatccgcgc caagagaaag cgggaggagc tgagcaacgt 2581 gctggcagcc atgaggaagg cggcggccaa gaaggattga gcggccgcga aggatctgcg 2641 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 2701 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa 2761 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 2821 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc 2881 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac 2941 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg 3001 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc 3061 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct 3121 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag 3181 acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca 3241 ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc 3301 agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc 3361 tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg 3421 agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt 3481 acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga 3541 tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca 3601 agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg 3661 tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg 3721 acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca 3781 tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt 3841 accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt 3901 ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc agaggaagtc 3961 ttctaacatg cggtgacgtg gaggagaatc ccggccctgc caccatggct gaacaagatg 4021 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 4081 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 4141 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 4201 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 4261 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4321 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4381 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4441 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4501 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4561 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4621 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4681 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4741 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4801 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 4861 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 4921 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 4981 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 5041 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 5101 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 5161 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 5221 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 5281 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 5341 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcggtacc 5401 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 5461 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg 5521 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 5581 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 5641 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 5701 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 5761 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 5821 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 5881 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc 5941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6061 gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 6121 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 6181 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 6241 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 6301 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 6361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 6421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 6481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6661 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 6901 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 6961 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 7021 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 7081 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 7141 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 7201 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 7261 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 7321 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 7381 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 7441 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 7501 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 7561 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 7621 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 7681 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 7741 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 7801 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 7861 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 7921 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 7981 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 8041 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 8101 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 8161 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 8221 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 8281 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 8341 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 8401 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 8461 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta 8521 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg 8581 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg 8641 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt 8701 gccaagctg //