LOCUS V035699 9128 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035699 VERSION V035699 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..3231 /label="SIRT5(NM_012241)" /note="SIRT5(NM_012241)" /gene="SIRT5" CDS 3238..3264 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3268..3294 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3298..3324 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3331..4050 /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" promoter 4098..4309 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4322..4590 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4617..5216 /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 5223..5811 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5885..6118 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6190..6311 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6330..6465 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6503..6519) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6527..6543 /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(6551..6581) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6596..6617 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6905..7493) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7664..8524) /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(8525..8629) /gene="bla" /label="AmpR promoter" primer_bind 9103..9119 /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 catgcgacct ctccagattg tcccaagtcg attgatttcc 2341 cagctatatt gtggcctgaa gcctccagcg tccacacgaa accagatttg cctgaaaatg 2401 gctcggccaa gttcaagtat ggcagatttt cgaaagtttt ttgcaaaagc aaagcacata 2461 gtcatcatct caggagctgg tgttagtgca gaaagtggtg ttccgacctt cagaggagct 2521 ggaggttatt ggagaaaatg gcaagcccag gacctggcga ctcccctggc ctttgcccac 2581 aacccgtccc gggtgtggga gttctaccac taccggcggg aggtcatggg gagcaaggag 2641 cccaacgccg ggcaccgcgc catagccgag tgtgagaccc ggctgggcaa gcagggccgg 2701 cgagtcgtgg tcatcaccca gaacatcgat gagctgcacc gcaaggctgg caccaagaac 2761 cttctggaga tccatggtag cttatttaaa actcgatgta cctcttgtgg agttgtggct 2821 gagaattaca agagtccaat ttgtccagct ttatcaggaa aaggtgctcc agaacctgga 2881 actcaagatg ccagcatccc agttgagaaa cttccccggt gtgaagaggc aggctgcggg 2941 ggcttgctgc gacctcacgt cgtgtggttt ggagaaaacc tggatcctgc cattctggag 3001 gaggttgaca gagagctcgc ccactgtgat ttatgtctag tggtgggcac ttcctctgtg 3061 gtgtacccag cagccatgtt tgccccccag gtggctgcca ggggcgtgcc agtggctgaa 3121 tttaacacgg agaccacccc agctacgaac agattcaggt ttcatttcca gggaccctgt 3181 ggaacgactc ttcctgaagc ccttgcctgt catgaaaatg aaactgtttc tggatcctac 3241 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat 3301 ccttatgatg tcccagacta cgcagccacc atggtgagca agggcgagga gctgttcacc 3361 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg 3421 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 3481 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag 3541 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc 3601 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc 3661 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac 3721 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac 3781 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac 3841 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc 3901 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa 3961 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc 4021 actctcggca tggacgagct gtacaagtag ggatccgcgg ccgcgaagga tctgcgatcg 4081 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 4141 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 4201 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 4261 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg 4321 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg 4381 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta 4441 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc 4501 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt 4561 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga 4621 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 4681 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 4741 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 4801 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 4861 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 4921 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 4981 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 5041 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 5101 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 5161 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg 5221 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5281 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5341 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5401 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5461 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5521 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5581 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 5641 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5701 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5761 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct 5821 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 5881 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 5941 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6001 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6061 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6121 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6181 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6241 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6301 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 6361 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 6421 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 6481 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6541 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 6601 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 6661 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6721 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6781 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 6841 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 6901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 6961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7321 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 7381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 7981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8641 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 8701 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 8761 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 8821 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 8881 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 8941 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9001 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9061 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9121 ccaagctg //