LOCUS V021057 9787 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021057 VERSION V021057 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2826..3464 /label="FAM177A1(NM_001079519)" /note="FAM177A1(NM_001079519)" /gene="FAM177A1" regulatory 3495..3504 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3504..3545 /codon_start=1 /product="epitope tag from simian virus 5" /transl_table=1 /translation="GKPIPNPLLGLDST" /label="V5 tag" CDS 3552..4508 /codon_start=1 /product="mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" /transl_table=1 /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" /label="TurboID" CDS 4515..4541 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /transl_table=1 /translation="LPPLERLTL" /label="NES" promoter 4575..5074 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5095..5694 /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 5708..6296 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6503..7136 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7265..7281) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7289..7305 /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(7313..7343) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7358..7379 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7667..8255) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8426..9286) /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(9287..9391) /gene="bla" /label="AmpR promoter" polyA_signal 9439..9573 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ccaccatgga ccaggagcca gtgggcggtg tggaacgagg agaagccgtc gcagcctcgg 2881 gagctgcggc cgccgcggca ttcggggaat ctgcagggca gatgagtaac gaaagaggct 2941 ttgaaaatgt agaactggga gtcataggaa aaaagaagaa agtcccaagg agagtcatcc 3001 actttgttag tggtgaaaca atggaagaat atagcacaga tgaagacgaa gttgatggcc 3061 tggagaagaa agatgttttg cctactgttg atccgacaaa acttacctgg ggtccctact 3121 tatggtttta catgcttcgg gctgctacat caactctctc agtgtgtgac ttccttggag 3181 agaagattgc atctgttttg ggtatcagca ccccaaagta ccaatatgcc attgatgaat 3241 attatcggat gaagaaggag gaagaagaag aagaagaaga aaacaggatg tctgaagaag 3301 cagaaaaaca atatcaacag aataaattgc agactgattc cattgttcag acagatcaac 3361 cagagacagt gatatccagc tcatttgtga atgtcaattt tgaaatggag ggagacagtg 3421 aagtaattat ggaaagcaag caaaatccag tctctgtccc accaggtggt tctggtggtg 3481 gttctggtga attcgccacc atgggcaagc ccatccccaa ccccctgctg ggcctggaca 3541 gcaccgctag caaagacaat actgtgcctc tgaagctgat cgctctcctg gctaatggcg 3601 agttccatag tggcgaacag ctgggagaaa ccctgggcat gtccagggcc gctatcaaca 3661 agcacattca gactctgcgc gactggggcg tggacgtgtt caccgtgccc ggaaagggct 3721 actctctgcc cgagcctatc ccgctgctga acgctaaaca gattctggga cagctggacg 3781 gcgggagcgt ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa 3841 tcggcgagct gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa 3901 gcagagggag gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc 3961 gcctgaagcg gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag 4021 aagcgctgcg aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc 4081 tgcaggatag aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg 4141 cacagatcgt cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg 4201 tcaatcaggg ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg 4261 ccgctacgct gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg 4321 ctccatatct gccacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga 4381 tcattgggga caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc 4441 tgctggaaca ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg 4501 ccgaaaagct gcagctgcct cccctggagc gcctgaccct ggactaggat cccgcgactc 4561 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 4621 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 4681 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 4741 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 4801 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 4861 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 4921 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 4981 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 5041 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 5101 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 5161 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5221 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5281 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5341 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5401 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5461 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5521 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5581 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5641 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt 5701 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5761 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5821 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5881 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5941 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 6001 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 6061 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 6121 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 6181 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6241 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6301 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 6361 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6421 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6481 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 6541 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 6601 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 6661 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 6721 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 6781 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 6841 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 6901 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 6961 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 7021 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 7081 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 7141 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 7201 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat 7261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 7321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 7381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 7441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 7501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 7561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 7621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 7681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 8041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 8101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 8161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 8221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 8281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 8341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 8401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 8461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 8521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 8581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 9001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 9061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 9121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 9181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 9241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 9301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 9361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 9421 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 9481 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 9541 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 9601 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 9661 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9721 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9781 tagtagt //