LOCUS V044568 9265 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044568 VERSION V044568 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..3918 /label="FBXL6(NM_012162)" /note="FBXL6(NM_012162)" /gene="FBXL6" CDS 3925..3990 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4034..4245 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4258..4526 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4559..5353 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5360..5948 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6022..6255 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6327..6448 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6467..6602 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6640..6656) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6664..6680 /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(6688..6718) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6733..6754 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7042..7630) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7801..8661) /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(8662..8766) /gene="bla" /label="AmpR promoter" primer_bind 9240..9256 /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 catggctgcc ccagcctccc ggcaggtccg acgcagagcc 2341 cgggcagcgc cgcggccccg ctcggccgag gactggtggt gggaccggct ggcgccgagg 2401 ggctcggggt accacctgct gcagtccgac agcatgctgc tggtgctgtc cgaacccggc 2461 cccgcccggc cccgcgcaca gcggcgcgct tcccgccgca ctccccggca gccgccccgg 2521 ggccccagcg ccgcggccaa gcccaaggcc gggctcaggt ccgaggcggc ggccgcgccc 2581 gcacccgcac cggcacccac gcccacgccc gaggaagggc ccgacgcggg ctggggagac 2641 cgcattccct tggaaatcct ggtgcagatt ttcgggttgt tggtggcggc ggacggcccc 2701 atgcccttcc tgggcagggc tgcgcgcgtg tgccgccgct ggcaggaggc cgcttcccaa 2761 cccgcgctct ggcacaccgt gaccctgtcg tccccgctgg tcggccggcc tgccaagggc 2821 ggggtcaagg cggagaagaa gctccttgct tccctggagt ggcttatgcc caatcggttt 2881 tcacagctcc agaggctgac cctcatccac tggaagtctc aggtacaccc cgtgttgaag 2941 ctggtaggtg agtgctgtcc tcggctcact ttcctcaagc tctccggctg ccacggtgtg 3001 actgctgacg ctctggtcat gctagccaaa gcctgctgcc agctccatag cctggaccta 3061 cagcactcca tggtggagtc cacagctgtg gtgagcttct tggaggaggc agggtcccga 3121 atgcgcaagt tgtggctgac ctacagctcc cagacgacag ccatcctggg cgcactgctg 3181 ggcagctgct gcccccagct ccaggtcctg gaggtgagca ccggcatcaa ccgtaatagc 3241 attccccttc agctgcctgt cgaggctctg cagaaaggct gccctcagct ccaggtgctg 3301 cggctgttga acctgatgtg gctgcccaag cctccgggac gaggggtggc tcccggacca 3361 ggcttcccta gcctagagga gctctgcctg gcgagctcaa cctgcaactt tgtgagcaac 3421 gaggtcctgg gccgcctact ccacggctct cccaacctgc gcttactgga tcttcgtggc 3481 tgtgcgcgca tcacgccggc tggccttcag gatctgccat gtcgggagct ggagcagctt 3541 catctgggcc tgtatggcac gtcagaccgg ctgactctag ccaaggaggg cagccccttt 3601 ttgacccaga agtggtgcca tacactgcga gaactggact tgagtggcca ggggttcagt 3661 gagaaggacc tggagcaggc cctggctgcc ttcttaagca cccctggggg ctcacaccca 3721 gccctgtgct ctcttaacct caggggcacc cgggtcacac caagcactgt cagctctgtg 3781 atcagcggct gcccgggcct gctctacctc aacctggagt cctgccgctg ccttccccgg 3841 ggtctgaagc gggcctaccg gggcctggag gaagtccagt ggtgtctgga gcagctgctc 3901 accagcccct cacccagcgg atccgactac aaagaccatg acggtgatta taaagatcat 3961 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 4021 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4081 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4141 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4201 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4261 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4321 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4381 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4441 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4501 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct aggccaccat 4561 ggctgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 4621 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 4681 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 4741 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 4801 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 4861 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 4921 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 4981 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5041 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 5101 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5161 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5221 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5281 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5341 cgagttcttc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 5401 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 5461 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 5521 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 5581 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 5641 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 5701 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 5761 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 5821 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 5881 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 5941 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 6001 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc 6061 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 6121 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 6181 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 6241 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 6301 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 6361 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 6421 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 6481 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 6541 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 6601 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg 6661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 6781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 6841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7441 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8761 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 8821 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 8881 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 8941 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 9061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 9121 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 9181 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 9241 taaaacgacg gccagtgcca agctg //