LOCUS V033350 8945 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033350 VERSION V033350 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..3039 /label="Crisp3(NM_009639)" /note="Crisp3(NM_009639)" /gene="Crisp3" promoter 3104..3315 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3328..3596 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3625..3634 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3655..4317 /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 4387..4440 /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 4441..5040 /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 5041..5628 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5702..5935 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6007..6128 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6147..6282 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6320..6336) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6344..6360 /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(6368..6398) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6413..6434 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6722..7310) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7481..8341) /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(8342..8446) /gene="bla" /label="AmpR promoter" primer_bind 8920..8936 /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 cattaatgct tgtgctgttc 2341 ttcctggctg ctgtactgcc cccatccctt cttcaagata actctcagga gaacagtctt 2401 gagaaacttt caaccagtaa aaaatcagtc caagaagaga ttgtaagcaa gcacaaccaa 2461 ttgagacgaa aggtttctcc atctggcagt gacttactaa atatggaatg gaactatgat 2521 gctcaagtga atgctcagca acgggcagac aagtgtacat tcagtcacag tcctatagaa 2581 ctcaggacaa ctaatttaaa atgtggtgag aatttgttca tgtcatctta ccttgtacca 2641 tggtcttctg taatccaagg atggtataat gaatccaaag gtcttatatt tggtgtgggc 2701 ccaaagcaaa atgttagtgt ggttggacat catactcagg ttgtttggaa atcaaattta 2761 caagttgcat gtggagttgc tgaatgccct gaaaatccac tgagatactt ttatgtttgt 2821 cgctattgtc ctgtattgaa ttacagtggc cattatccaa gcaggccata cctagcttac 2881 acagcaagag caccatgtgc cagttgtcct gatcgctgtg aagatggact gtgcaccaag 2941 agttgtcaat ataaggatat gtctttttgg tgtaaacgtc tggaatacgt ctgtaaacat 3001 ccaggtctta aaaaacgttg cctagctaca tgccaatgtt aaggatccct gcgatatcca 3061 gcgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3121 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3181 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3241 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3301 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 3361 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 3421 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 3481 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 3541 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 3601 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 3661 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 3721 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 3781 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 3841 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 3901 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 3961 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4021 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4081 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4141 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4201 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4261 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4321 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 4381 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 4441 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 4501 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 4561 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 4621 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 4681 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 4741 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 4801 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 4861 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 4921 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 4981 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5041 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5101 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5161 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5221 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5281 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5341 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5401 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 5461 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 5521 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 5581 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 5641 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5701 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 5761 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5821 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 5881 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 5941 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6001 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6061 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6121 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6181 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6241 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6301 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6361 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6421 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6481 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 6961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7141 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 7921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 7981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8461 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8521 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 8581 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 8641 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 8701 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8761 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 8821 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 8881 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 8941 agctg //