LOCUS V044825 9118 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044825 VERSION V044825 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" enhancer 1933..2236 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2237..2440 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2475..4337 /label="KLHL40(NM_152393)" /note="KLHL40(NM_152393)" /gene="KLHL40" CDS 4344..4409 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 4419..4475 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 4488..5207 /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" misc_feature 5214..5801 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5875..6108 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6180..6301 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6320..6455 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6493..6509) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6517..6533 /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(6541..6571) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6586..6607 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6895..7483) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7654..8514) /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(8515..8619) /gene="bla" /label="AmpR promoter" primer_bind 9093..9109 /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 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 1981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 2041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 2101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 2161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 2221 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 2281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 2341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 2401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat 2461 ccgctagcgc caccatggcg ctgggcttgg agcaggcgga ggagcagcgg ttgtaccagc 2521 agacgctcct gcaagacggg ctcaaagaca tgctggacca tggcaagttc ctcgactgtg 2581 tggtgcgggc gggcgagcgc gagttcccgt gccatcgcct ggtgctggcc gcctgcagcc 2641 cctacttccg ggcgcgcttt ctagccgagc cggagcgcgc gggcgagctg cacctggagg 2701 aggtgtcccc ggacgtggtg gcccaggtgc tgcactacct gtacacatca gagatcgcgc 2761 tggatgaggc gagcgtgcag gatttgttcg ccgcggcaca ccgcttccag atcccttcca 2821 tcttcaccat ctgcgtgtcc ttcctgcaga agcgcctgtg cctctccaac tgcttggccg 2881 tcttccgtct cggcctcctg ctcgactgcg cgcgtctcgc cgtggctgcc cgcgacttca 2941 tctgcgctca cttcacgctg gtggcgcgcg acgctgactt cctcggactc tcggccgacg 3001 agctcatcgc catcatctcc agcgacggcc ttaacgtgga gaaggaggag gcagtgttcg 3061 aggcggtgat gcggtgggcg ggtagcggcg acgccgaggc gcaggctgag cgccagcgcg 3121 cgctgcccac cgtcttcgag agcgtgcgct gccgcttgct gccgcgcgcc tttctggaaa 3181 gccgcgtgga gcgccaccct ctcgtgcgtg cccagcccga gttgctgcgc aaggtgcaga 3241 tggtgaagga tgcacacgag ggccgcatca ccacgctgcg gaagaaaaag aaggggaagg 3301 atggagccgg ggccaaggag gctgataagg gcacaagcaa agccaaagca gaggaggatg 3361 aggaggccga acgtatcctt cctgggatcc tcaatgacac cctgcgcttc ggcatgttcc 3421 tgcaggatct catcttcatg atcagtgagg agggcgctgt ggcctacgat ccagcagcca 3481 acgagtgcta ctgtgcttcc ctctccaacc aggtccccaa gaaccacgtc agcctggtta 3541 ccaaggagaa ccaggtcttc gtggctggag gcctcttcta caacgaagac aacaaagagg 3601 accccatgag cgcatacttc ctgcagtttg accatctgga ctcagagtgg ctggggatgc 3661 caccgctgcc ctcgccccgc tgcctctttg gcctgggaga agctctcaac tccatctacg 3721 tggtcggtgg cagagagatc aaggacggcg agcgctgcct ggactcggtc atgtgctacg 3781 acaggctgtc attcaaatgg ggtgaatcgg acccgctgcc ttacgtggtg tatggccaca 3841 cagtgctctc ccacatggac cttgtctacg taattggcgg caaaggcagt gacaggaagt 3901 gcctgaacaa gatgtgcgtc tatgacccca agaagtttga gtggaaggag ctggcaccca 3961 tgcagaccgc ccgctcactc tttggggcca ctgtccatga tggccgcatt atcgtggcag 4021 ctggggtcac cgacacaggg ctgaccagtt ctgccgaagt gtacagcatc acagacaaca 4081 agtgggcacc cttcgaggcc ttcccacagg agcgtagctc actcagcctg gtcagcctgg 4141 tgggtaccct ctatgccatt ggtggctttg ccacactgga gacggagtct ggagagctgg 4201 ttcccacaga gctcaatgac atctggaggt ataacgagga ggagaagaaa tgggagggtg 4261 tcctgcggga gatcgcctat gcagcaggtg ccaccttcct accagtgcgg ctcaatgtgc 4321 tgtgcctgac taagatggtc gacgactaca aagaccatga cggtgattat aaagatcatg 4381 acatcgacta caaggatgac gatgacaagg gaagcggagc cactaacttc tccctgttga 4441 aacaagcagg ggatgtcgaa gagaatcccg ggccagaatt cgccaccatg gtgagcaagg 4501 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg 4561 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 4621 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc 4681 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct 4741 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg 4801 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg 4861 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca 4921 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga 4981 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc 5041 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc 5101 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 5161 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagga tccaatcaac 5221 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 5281 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 5341 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 5401 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 5461 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 5521 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 5581 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg 5641 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 5701 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc 5761 gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct ttaagaccaa 5821 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 5881 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 5941 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 6001 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 6061 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 6121 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 6181 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 6241 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6301 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 6361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 6421 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 6481 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6541 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6601 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6661 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6721 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6781 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6841 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6901 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6961 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7021 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7081 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7141 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7201 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7261 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7321 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7381 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7441 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7501 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7561 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7621 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7681 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7741 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7801 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7861 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7921 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7981 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8041 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8101 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8161 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8221 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8281 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8341 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8401 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8461 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8521 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8581 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8641 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8701 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8761 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8821 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8881 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8941 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9001 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9061 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //