LOCUS V041324 9665 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041324 VERSION V041324 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3018 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 3055..4584 /label="LMAN1(NM_005570)" /note="LMAN1(NM_005570)" /gene="LMAN1" promoter 4635..4846 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4859..5127 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5154..5753 /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 5760..6348 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6422..6655 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6727..6848 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6867..7002 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7040..7056) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7064..7080 /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(7088..7118) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7133..7154 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7442..8030) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8201..9061) /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(9062..9166) /gene="bla" /label="AmpR promoter" primer_bind 9640..9656 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg 2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggcg 3061 ggatccaggc aaaggggtct ccgggccaga gttcggccgc tgttctgcgc cttgctgctg 3121 tcactcggtc gcttcgtccg gggcgacggc gtgggaggag accccgcggt cgcgttgcca 3181 catcgccgtt tcgagtacaa atacagcttc aaggggccgc acctggtgca gagcgacggg 3241 accgtgccct tctgggccca cgcggggaat gctattccaa gttcagatca aattcgagta 3301 gcaccatctt taaaaagcca aagaggctca gtgtggacaa agacaaaagc ggcctttgag 3361 aactgggaag ttgaggtgac atttcgagtg actggaagag gtcgaattgg agctgatggc 3421 ctagcaattt ggtatgcaga aaatcaaggc ttggagggcc ctgtgtttgg atcagctgat 3481 ctgtggaatg gtgttggaat attttttgat tcttttgaca atgatggaaa gaaaaataat 3541 cctgctatag taattatagg caacaatgga caaatccatt atgaccatca aaatgacggg 3601 gctagtcaag ctttggcaag ttgccagagg gacttccgca acaaacccta tcctgtccga 3661 gcaaagatta cctattacca gaacacactg acagtaatga tcaataatgg ctttacacca 3721 gataaaaatg attatgaatt ttgtgccaaa gtggaaaata tgattatccc tgcacaaggg 3781 cattttggaa tatctgctgc aactggaggt cttgcagatg accatgatgt cctttctttt 3841 ctgactttcc agttgactga acctggaaaa gagccgccca caccagataa agaaatttcg 3901 gaaaaggaaa aagaaaagta tcaggaggaa tttgagcact ttcaacaaga attggataaa 3961 aaaaaagagg aattccagaa gggccacccc gacctccaag ggcagcctgc ggaggaaata 4021 tttgagagtg taggagatcg agagctaaga caagtctttg aaggacagaa tcgtattcat 4081 cttgaaatca agcagctgaa ccggcagtta gatatgattc ttgatgaaca gagaagatat 4141 gtctcttcct taacagagga aatctctaaa agaggagcag gaatgcctgg gcagcatggg 4201 cagattactc aacaagaact ggatactgtt gtgaaaactc agcatgagat tctgagacaa 4261 gtaaatgaaa tgaaaaattc catgagtgaa accgtcagac tggtcagtgg aatgcagcac 4321 cctggctctg ctggaggcgt ctatgagaca acacagcact tcattgacat caaagagcac 4381 ctgcacatag taaagaggga catagataac ttagtgcagc gaaatatgcc atcaaatgaa 4441 aagccgaaat gcccagaact accaccattt ccatcatgtt tgtctacggt ccacttcatt 4501 atatttgttg tggtgcaaac tgtattattc attggttata tcatgtatag gtctcagcaa 4561 gaagcagctg ccaaaaaatt cttttgactc gaggcggccg cgaaggatct gcgatcgctc 4621 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 4681 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 4741 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 4801 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 4861 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 4921 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 4981 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 5041 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 5101 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 5161 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 5221 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 5281 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5341 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5401 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 5461 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 5521 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 5581 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5641 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5701 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 5761 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5821 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5881 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5941 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 6001 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6061 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6121 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 6181 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6241 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 6301 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 6361 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6421 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6481 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6541 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6601 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6661 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6721 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6841 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6901 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6961 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7021 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7861 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9181 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9241 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9301 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9361 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9421 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9481 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9541 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9601 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9661 agctg //