LOCUS V038281 9673 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038281 VERSION V038281 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 3049..4722 /label="Atl1(NM_001009831)" /note="Atl1(NM_001009831)" /gene="Atl1" CDS 4729..4794 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4838..5049 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5062..5330 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5363..5761 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 5768..6356 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6430..6663 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6735..6856 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6875..7010 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7048..7064) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7072..7088 /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(7096..7126) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7141..7162 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7450..8038) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8209..9069) /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(9070..9174) /gene="bla" /label="AmpR promoter" primer_bind 9648..9664 /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 aggtggaggt agtggaggag gcggatcaat ggctaagagc 3061 cgcagggaca gaaacagttg gggtgggttt tcagagaagt caagtgattg gagctcagaa 3121 gaggaggagc cggtgagaaa ggcaggaccc gtccaagtcc tcatcgtcaa agacgaccat 3181 tcctttgagt tagatgaggc tgcactgaac cggatccttc tgtctgaggc cgtcagagac 3241 aaggaagtgg ttgccgtgtc tgttgctgga gcctttagaa aaggaaaatc cttcctgatg 3301 gacttcatgt tacggtacat gtacaaccag gaatctgttg attgggttgg agattacaat 3361 gaaccattga ctggattttc atggagaggt ggatctgaac gagaaaccac aggaatccag 3421 atctggagcg aagttttcct catcaataag cttgatggta aaaaggttgc agtgttactg 3481 atggatactc agggaacctt tgacagtcag tcaactttga gagattcggc cacagtattt 3541 gcccttagca caatgataag ctcaatacag gtctacaact tatcccaaaa tgtccaggag 3601 gacgaccttc agcatctcca gcttttcact gagtatggca ggttggcaat ggaagaaaca 3661 ttcctgaagc cttttcagag tctgatattt cttgttcgag actggagttt cccatacgaa 3721 ttctcatacg gagctgatgg tggcgccaag tttttggaaa aacgcctcaa ggtctcaggg 3781 aaccagcacg aagaactaca gaatgtcaga aagcacatcc attcctgttt taccaacatt 3841 tcctgttttc tcctacctca tcctggctta aaagtagcta cgaacccaaa ctttgatgga 3901 aaactaaaag aaatagatga cgaattcatc aaaaacttga aaatactgat tccgtggctc 3961 cttagtcccg agagcctaga tattaaagag atcaatggga ataaaatcac ctgccgaggt 4021 ctgctggagt acttcaaggc gtacataaag atctaccaag gggaagaatt gccacatccc 4081 aagtccatgt tacaggccac agcagaagcc aacaacctag cagctgttgc aaccgccaag 4141 gacacataca acaagaagat ggaggagatt tgtggtggtg acaaaccatt cttggccccc 4201 aatgacctgc aaaccaagca cctgcagcta aaagaagact ctgtgaagct gttccgaggt 4261 gtgaagaaga tgggtgggga agaattcagc cggcgctacc tacagcagct ggagagtgaa 4321 atagacgaac tctacatcca gtacatcaag cacaatgaca gcaaaaatat cttccatgcc 4381 gctcgcaccc cagccacact gtttgttgtc atctttatca cgtatgtcat cgctggtgtg 4441 actggattca ttggtttgga catcatagct agcctgtgca acatgatcat gggactgaca 4501 ctcatcacgc tgtgcacctg ggcatacatc cgctactctg gggagtatcg agagctgggc 4561 gctgtaatcg accaggtagc tgcagctttg tgggaccagg gaagcacaaa tgaggcttta 4621 tacaagctgt acagtgcagc agcaacccac agacacctgt atcagcaagc ctttcctgca 4681 cccaagtcag aacctactga gcaaccggaa aagaagaaaa ttggatccga ctacaaagac 4741 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 4801 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 4861 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 4921 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 4981 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 5041 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 5101 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 5161 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 5221 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 5281 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 5341 ccggcgccta cgctaggcca ccatggccaa gcctttgtct caagaagaat ccaccctcat 5401 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag 5461 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg 5521 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct 5581 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg 5641 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg 5701 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta 5761 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5821 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5881 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5941 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 6001 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 6061 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 6121 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 6181 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 6241 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6301 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6361 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6421 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 6481 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6541 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6601 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6661 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6721 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6841 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6901 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6961 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 7021 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 7081 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7141 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7201 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7261 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7321 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7381 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7861 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9181 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9241 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9301 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9361 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9421 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9481 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9541 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9601 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9661 cagtgccaag ctg //