LOCUS V045930 9358 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045930 VERSION V045930 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..3384 /label="STOML2(NM_013442)" /note="STOML2(NM_013442)" /gene="STOML2" CDS 3391..3456 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3500..3711 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3724..3992 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4021..4030 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4051..4713 /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 4783..4836 /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 4837..5436 /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 5437..6025 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6099..6332 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6404..6525 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6565..6700 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6733..6749) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6757..6773 /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(6781..6811) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6826..6847 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7135..7723) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7894..8754) /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(8755..8859) /gene="bla" /label="AmpR promoter" primer_bind 9333..9349 /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 gccaccatgc tggcgcgcgc ggcgcggggc 2341 actggggccc ttttgctgag gggctctcta ctggcttctg gccgcgctcc gcgccgcgcc 2401 tcctctggat tgccccgaaa caccgtggta ctgttcgtgc cgcagcagga ggcctgggtg 2461 gtggagcgaa tgggccgatt ccaccggatc ctggagcctg gtttgaacat cctcatccct 2521 gtgttagacc ggatccgata tgtgcagagt ctcaaggaaa ttgtcatcaa cgtgcctgag 2581 cagtcggctg tgactctcga caatgtaact ctgcaaatcg atggagtcct ttacctgcgc 2641 atcatggacc cttacaaggc aagctacggt gtggaggacc ctgagtatgc cgtcacccag 2701 ctagctcaaa caaccatgag atcagagctc ggcaaactct ctctggacaa agtcttccgg 2761 gaacgggagt ccctgaatgc cagcattgtg gatgccatca accaagctgc tgactgctgg 2821 ggtatccgct gcctccgtta tgagatcaag gatatccatg tgccaccccg ggtgaaagag 2881 tctatgcaga tgcaggtgga ggcagagcgg cggaaacggg ccacagttct agagtctgag 2941 gggacccgag agtcggccat caatgtggca gaagggaaga aacaggccca gatcctggcc 3001 tccgaagcag aaaaggctga acagataaat caggcagcag gagaggccag tgcagttctg 3061 gcgaaggcca aggctaaagc tgaagctatt cgaatcctgg ctgcagctct gacacaacat 3121 aatggagatg cagcagcttc actgactgtg gccgagcagt atgtcagcgc gttctccaaa 3181 ctggccaagg actccaacac tatcctactg ccctccaacc ctggcgatgt caccagcatg 3241 gtggctcagg ccatgggtgt atatggagcc ctcaccaaag ccccagtgcc agggactcca 3301 gactcactct ccagtgggag cagcagagat gtccagggta cagatgcaag tcttgatgag 3361 gaacttgatc gagtcaagat gagtggatcc gactacaaag accatgacgg tgattataaa 3421 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3481 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3541 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3601 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3661 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3721 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3781 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3841 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3901 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3961 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4021 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4081 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4141 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4201 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4261 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4321 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4381 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4441 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4501 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4561 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4621 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4681 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4741 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4801 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4861 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4921 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4981 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5041 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5101 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5161 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5221 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5281 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5341 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5401 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5461 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5521 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5581 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5641 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5701 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5761 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5821 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5881 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5941 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6001 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6061 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6121 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6181 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6241 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6301 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6361 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6421 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6481 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6541 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6601 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6661 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6721 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6781 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6841 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7561 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8821 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8881 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8941 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9001 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9061 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9121 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9181 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9241 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9301 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //