LOCUS V032997 9617 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032997 VERSION V032997 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="cyan variant of GFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG IKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="CFP" CDS 3034..4536 /label="SLC7A11(NM_014331)" /note="SLC7A11(NM_014331)" /gene="SLC7A11" promoter 4587..4798 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4811..5079 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5106..5705 /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 5712..6300 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6374..6607 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6679..6800 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6819..6954 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6992..7008) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7016..7032 /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(7040..7070) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7085..7106 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7394..7982) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8153..9013) /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(9014..9118) /gene="bla" /label="AmpR promoter" primer_bind 9592..9608 /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 ggttcagcgt gtccggcgag 2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc 2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2581 gtccaggagc gtaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 2701 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc 2761 accgccgaca agcagaagaa cggcatcaag gcccacttca 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 aggaggggga agcatggtca gaaagcctgt tgtgtccacc 3061 atctccaaag gaggttacct gcagggaaat gttaacggga ggctgccttc cctgggcaac 3121 aaggagccac ctgggcagga gaaagtgcag ctgaagagga aagtcacttt actgagggga 3181 gtctccatta tcattggcac catcattgga gcaggaatct tcatctctcc taagggcgtg 3241 ctccagaaca cgggcagcgt gggcatgtct ctgaccatct ggacggtgtg tggggtcctg 3301 tcactatttg gagctttgtc ttatgctgaa ttgggaacaa ctataaagaa atctggaggt 3361 cattacacat atattttgga agtctttggt ccattaccag cttttgtacg agtctgggtg 3421 gaactcctca taatacgccc tgcagctact gctgtgatat ccctggcatt tggacgctac 3481 attctggaac cattttttat tcaatgtgaa atccctgaac ttgcgatcaa gctcattaca 3541 gctgtgggca taactgtagt gatggtccta aatagcatga gtgtcagctg gagcgcccgg 3601 atccagattt tcttaacctt ttgcaagctc acagcaattc tgataattat agtccctgga 3661 gttatgcagc taattaaagg tcaaacgcag aactttaaag acgccttttc aggaagagat 3721 tcaagtatta cgcggttgcc actggctttt tattatggaa tgtatgcata tgctggctgg 3781 ttttacctca actttgttac tgaagaagta gaaaaccctg aaaaaaccat tccccttgca 3841 atatgtatat ccatggccat tgtcaccatt ggctatgtgc tgacaaatgt ggcctacttt 3901 acgaccatta atgctgagga gctgctgctt tcaaatgcag tggcagtgac cttttctgag 3961 cggctactgg gaaatttctc attagcagtt ccgatctttg ttgccctctc ctgctttggc 4021 tccatgaacg gtggtgtgtt tgctgtctcc aggttattct atgttgcgtc tcgagagggt 4081 caccttccag aaatcctctc catgattcat gtccgcaagc acactcctct accagctgtt 4141 attgttttgc accctttgac aatgataatg ctcttctctg gagacctcga cagtcttttg 4201 aatttcctca gttttgccag gtggcttttt attgggctgg cagttgctgg gctgatttat 4261 cttcgataca aatgcccaga tatgcatcgt cctttcaagg tgccactgtt catcccagct 4321 ttgttttcct tcacatgcct cttcatggtt gccctttccc tctattcgga cccatttagt 4381 acagggattg gcttcgtcat cactctgact ggagtccctg cgtattatct ctttattata 4441 tgggacaaga aacccaggtg gtttagaata atgtcggaga aaataaccag aacattacaa 4501 ataatactgg aagttgtacc agaagaagat aagttatagg gatccgcggc cgcgaaggat 4561 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa 4621 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg 4681 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat 4741 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct 4801 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca 4861 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc 4921 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg 4981 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta 5041 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac 5101 gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg 5161 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc 5221 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc 5281 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc 5341 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga 5401 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca 5461 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc 5521 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct 5581 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca 5641 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg 5701 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 5761 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 5821 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 5881 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5941 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 6001 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 6061 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 6121 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 6181 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 6241 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 6301 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 6361 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct 6421 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 6481 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 6541 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 6601 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 6661 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat 6721 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 6781 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca 6841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 6901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact 6961 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7021 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7081 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7141 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7201 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7261 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7321 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7381 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 7441 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 7501 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 7561 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 7621 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 7681 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 7741 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 7801 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 7861 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 7921 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 7981 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 8041 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 8101 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8161 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8221 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8281 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8341 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 8401 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 8461 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 8521 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 8581 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 8641 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 8701 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 8761 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 8821 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 8881 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 8941 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 9001 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 9061 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9121 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 9181 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa 9241 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 9301 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 9361 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca 9421 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt 9481 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 9541 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 9601 cggccagtgc caagctg //