LOCUS V028028 9118 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028028 VERSION V028028 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3144 /label="METTL1(NM_005371)" /note="METTL1(NM_005371)" /gene="METTL1" CDS 3151..3216 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3260..3471 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3484..3752 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3781..3790 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3811..4473 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 4543..4596 /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 Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 4597..5196 /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 5197..5785 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5859..6092 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6164..6285 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6325..6460 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg cagccgagac tcggaacgtg 2341 gccggagcag aggccccacc gccccagaag cgctactacc ggcaacgtgc tcactccaac 2401 cccatggcgg accacacgct gcgctaccct gtgaagccag aggagatgga ctggtctgag 2461 ctatacccag agttcttcgc tccactcact caaaatcaga gccacgatga cccaaaggat 2521 aagaaagaaa agagagctca ggcccaagtg gagtttgcag acataggctg tggctatggt 2581 ggcctgttag tggaactgtc accgctgttc ccagacacac ttattctggg tctggagatc 2641 cgggtgaagg tctcagacta tgtacaagac cggattcggg ccctacgcgc agctcctgca 2701 ggtggcttcc agaacatcgc ctgtctccgt agcaatgcca tgaagcacct tcctaacttc 2761 ttctacaagg gccagctgac aaagatgttc ttcctcttcc ccgacccaca tttcaagcgg 2821 acaaagcaca agtggcgaat catcagtccc accctgctag cagaatatgc ctacgtgcta 2881 agagttgggg ggctggtgta taccataacc gatgtgctgg agctacacga ctggatgtgc 2941 actcatttcg aagagcaccc actgtttgag cgtgtgcctc tggaggacct gagtgaagac 3001 cccgttgtgg gacatctagg cacctcaact gaggagggga agaaagttct acgtaatgga 3061 gggaagaatt tcccagccat cttccgaaga atacaagatc ccgtcctcca ggcagtgacc 3121 tcccaaacca gcctgcctgg tcacggatcc gactacaaag accatgacgg tgattataaa 3181 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3241 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3301 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3361 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3421 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3481 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3541 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3601 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3661 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3721 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3781 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 3841 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 3901 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 3961 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4021 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4081 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4141 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4201 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4261 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4321 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4381 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4441 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4501 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4561 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4621 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4681 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4741 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4801 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 4861 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 4921 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 4981 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5041 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5101 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5161 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5221 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5281 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5341 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5401 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5461 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5521 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5581 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5641 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5701 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5761 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 5821 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 5881 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 5941 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6001 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6061 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6121 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6301 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6361 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6421 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6481 attcgtaatc atgtcatagc 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 //