LOCUS V027258 9544 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027258 VERSION V027258 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..3570 /label="SERPINA1(NM_000295)" /note="SERPINA1(NM_000295)" /gene="SERPINA1" CDS 3577..3642 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3686..3897 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3910..4178 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4207..4216 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4237..4899 /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 4969..5022 /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 5023..5622 /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 5623..6211 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6285..6518 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6590..6711 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6751..6886 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6919..6935) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6943..6959 /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(6967..6997) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7012..7033 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7321..7909) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8080..8940) /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(8941..9045) /gene="bla" /label="AmpR promoter" primer_bind 9519..9535 /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 cgtcttctgt ctcgtggggc 2341 atcctcctgc tggcaggcct gtgctgcctg gtccctgtct ccctggctga ggatccccag 2401 ggagatgctg cccagaagac agatacatcc caccatgatc aggatcaccc aaccttcaac 2461 aagatcaccc ccaacctggc tgagttcgcc ttcagcctat accgccagct ggcacaccag 2521 tccaacagca ccaatatctt cttctcccca gtgagcatcg ctacagcctt tgcaatgctc 2581 tccctgggga ccaaggctga cactcacgat gaaatcctgg agggcctgaa tttcaacctc 2641 acggagattc cggaggctca gatccatgaa ggcttccagg aactcctccg taccctcaac 2701 cagccagaca gccagctcca gctgaccacc ggcaatggcc tgttcctcag cgagggcctg 2761 aagctagtgg ataagttttt ggaggatgtt aaaaagttgt accactcaga agccttcact 2821 gtcaacttcg gggacaccga agaggccaag aaacagatca acgattacgt ggagaagggt 2881 actcaaggga aaattgtgga tttggtcaag gagcttgaca gagacacagt ttttgctctg 2941 gtgaattaca tcttctttaa aggcaaatgg gagagaccct ttgaagtcaa ggacaccgag 3001 gaagaggact tccacgtgga ccaggtgacc accgtgaagg tgcctatgat gaagcgttta 3061 ggcatgttta acatccagca ctgtaagaag ctgtccagct gggtgctgct gatgaaatac 3121 ctgggcaatg ccaccgccat cttcttcctg cctgatgagg ggaaactaca gcacctggaa 3181 aatgaactca cccacgatat catcaccaag ttcctggaaa atgaagacag aaggtctgcc 3241 agcttacatt tacccaaact gtccattact ggaacctatg atctgaagag cgtcctgggt 3301 caactgggca tcactaaggt cttcagcaat ggggctgacc tctccggggt cacagaggag 3361 gcacccctga agctctccaa ggccgtgcat aaggctgtgc tgaccatcga cgagaaaggg 3421 actgaagctg ctggggccat gtttttagag gccataccca tgtctatccc ccccgaggtc 3481 aagttcaaca aaccctttgt cttcttaatg attgaacaaa ataccaagtc tcccctcttc 3541 atgggaaaag tggtgaatcc cacccaaaaa ggatccgact acaaagacca tgacggtgat 3601 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3661 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3721 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3781 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3841 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3901 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3961 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4021 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4081 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4141 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4201 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4261 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4321 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4381 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4441 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4501 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4561 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4621 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4681 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4741 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4801 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4861 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4921 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4981 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5041 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5101 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5161 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5221 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5281 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5341 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5401 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5461 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5521 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5581 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5641 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5701 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5761 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5821 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5881 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5941 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6001 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6061 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6121 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6181 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6241 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6301 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6361 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6421 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6481 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6541 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6601 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6661 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6721 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6901 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6961 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7021 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7141 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7201 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7261 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7741 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7861 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7921 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7981 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8161 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8221 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8281 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8341 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8401 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8461 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8521 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8581 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8641 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8701 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8761 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8821 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8881 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8941 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9001 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9061 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9121 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9181 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9241 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9301 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9361 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9421 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9481 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9541 gctg //