LOCUS V044411 8758 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044411 VERSION V044411 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..2784 /label="RPL23A(NM_000984)" /note="RPL23A(NM_000984)" /gene="RPL23A" CDS 2791..2856 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2900..3111 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3124..3392 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3421..3430 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3451..4113 /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 4183..4236 /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 4237..4836 /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 4837..5425 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5499..5732 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5804..5925 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5965..6100 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6133..6149) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6157..6173 /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(6181..6211) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6226..6247 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6535..7123) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7294..8154) /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(8155..8259) /gene="bla" /label="AmpR promoter" primer_bind 8733..8749 /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 cgccgaaagc gaagaaggaa 2341 gctcctgccc ctcctaaagc tgaagccaaa gcgaaggctt taaaggccaa gaaggcagtg 2401 ttgaaaggtg tccacagcca caaaaagaag aagatccgca cgtcacccac cttccggcgg 2461 ccgaagacac tgcgactccg gagacagccc aaatatcctc ggaagagcgc tcccaggaga 2521 aacaagcttg accactatgc tatcatcaag tttccgctga ccactgagtc tgccatgaag 2581 aagatagaag acaacaacac acttgtgttc attgtggatg ttaaagccaa caagcaccag 2641 attaaacagg ctgtgaagaa gctgtatgac attgatgtgg ccaaggtcaa caccctgatt 2701 cggcctgatg gagagaagaa ggcatatgtt cgactggctc ctgattacga tgctttggat 2761 gttgccaaca aaattgggat catcggatcc gactacaaag accatgacgg tgattataaa 2821 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 2881 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 2941 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3001 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3061 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3121 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3181 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3241 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3301 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3361 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3421 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 3481 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 3541 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 3601 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 3661 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 3721 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 3781 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 3841 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 3901 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 3961 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4021 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4081 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4141 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4201 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4261 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4321 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4381 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4441 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 4501 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 4561 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 4621 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 4681 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 4741 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 4801 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 4861 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 4921 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 4981 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5041 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5101 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5161 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5221 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5281 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5341 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5401 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 5461 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 5521 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 5581 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 5641 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 5701 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 5761 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 5821 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 5881 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 5941 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6061 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6121 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6961 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8281 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8341 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8401 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8461 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8521 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8581 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8641 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8701 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //