LOCUS V029521 9365 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029521 VERSION V029521 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..4117 /label="RPN1(NM_002950)" /note="RPN1(NM_002950)" /gene="RPN1" promoter 4168..4379 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4392..4660 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4689..4698 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4719..5381 /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" misc_feature 5460..6048 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6122..6355 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6427..6548 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6567..6702 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6740..6756) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6764..6780 /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(6788..6818) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6833..6854 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7142..7730) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7901..8761) /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(8762..8866) /gene="bla" /label="AmpR promoter" primer_bind 9340..9356 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgg aggcgccagc cgccggcttg tttctgctcc tgttgcttgg 2341 gacttgggcc ccggcgccgg gcagcgcctc ctccgaggca ccgccgctga tcaatgagga 2401 cgtgaagcgc acagtggacc taagcagcca cctggctaag gtgacggccg aggtggtcct 2461 ggcgcacctg ggcggcggct ccacgtcccg agctacctct ttcctgctgg ctttggagcc 2521 tgagctcgag gcccggctgg cgcacctggg cgtgcaggta aagggagaag atgaggaaga 2581 gaacaatttg gaagtacgtg aaaccaaaat taagggtaaa agtgggagat tcttcacagt 2641 caagctccca gttgctcttg atcctggggc caagatttca gtcattgtgg aaacagtcta 2701 cacccatgtg cttcatccgt atccaaccca gatcacccag tcagagaaac agtttgtggt 2761 gtttgagggg aaccattatt tctactctcc ctatccaacg aagacacaaa ccatgcgtgt 2821 gaagcttgcc tctcgaaatg tggagagcta caccaagctg gggaacccca cgcgctctga 2881 ggacctactg gattatgggc ctttcagaga tgtgcctgcc tatagtcagg atacttttaa 2941 agtacattat gagaacaaca gccctttcct gaccatcacc agcatgaccc gagtcattga 3001 agtctctcac tggggtaata ttgctgtgga agaaaatgtg gacttaaagc acacaggagc 3061 tgtgcttaag gggcctttct cacgctatga ttaccagaga cagccagata gtggaatatc 3121 ctccatccgt tcttttaaga ccatccttcc tgctgctgcc caggatgttt attaccggga 3181 tgagattggc aatgtttcta ccagccacct ccttattttg gatgactctg tagagatgga 3241 aatccggcct cgcttccctc tctttggcgg gtggaagacc cattacatcg ttggctacaa 3301 cctcccaagc tatgagtacc tctataattt gggtgaccag tatgcactga agatgaggtt 3361 tgtggaccat gtgtttgatg aacaagtgat agattctctg actgtgaaga tcatcctgcc 3421 tgaaggagcc aagaacattg aaattgatag tccctatgaa atcagccgtg ccccagatga 3481 gctgcactac acctatctgg acacatttgg ccgccctgtg attgttgcct acaagaaaaa 3541 tctggtagaa cagcacattc aggacattgt ggtccactac acgttcaaca aggtgctcat 3601 gctgcaggag cccctgctgg tggtggcggc cttctacatc ctgttcttca ccgttatcat 3661 ctatgttcgg ctggacttct ccatcaccaa ggatccagcc gcagaagcca ggatgaaggt 3721 agcctgcatc acagagcagg tcttgaccct ggtcaacaag agaataggcc tttaccgtca 3781 ctttgacgag accgtcaata ggtacaagca atcccgggac atctccaccc tcaacagtgg 3841 caagaagagc ctggagactg aacacaaggc cttgaccagt gagattgcac tgctgcagtc 3901 caggctgaag acagagggct ctgatctgtg cgacagagtg agcgaaatgc agaagctgga 3961 tgcacaggtc aaggagctgg tgctgaagtc ggcggtggag gctgagcgcc tggtggctgg 4021 caagctcaag aaagacacgt acattgagaa tgagaagctc atctcaggaa agcgccagga 4081 gctggtcacc aagatcgacc acatcctgga tgccctgtag ggatccgcgg ccgcgaagga 4141 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga 4201 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact 4261 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata 4321 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc 4381 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc 4441 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt 4501 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt 4561 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct 4621 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta 4681 cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag atcgagtgcc 4741 gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga gagggcaccc 4801 ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg accttcagcc 4861 cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc taccccagcg 4921 gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac acccgcatcg 4981 agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac gaggccggcc 5041 gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc gtgatcttca 5101 ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg ggcgataacg 5161 tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac tacagcttcg 5221 tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg cagaacgggg 5281 gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag ctgggcatcg 5341 tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc gctcagtcgt 5401 ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc taagtcgaca 5461 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5521 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5581 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5641 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 5701 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 5761 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5821 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 5881 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5941 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 6001 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 6061 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6121 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6181 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6241 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6301 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6361 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6421 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6541 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6601 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6661 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6721 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7561 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8881 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8941 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9001 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9061 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9121 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9181 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9241 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9301 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9361 agctg //