LOCUS V027747 9625 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027747 VERSION V027747 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..3651 /label="SLC52A2(NM_024531)" /note="SLC52A2(NM_024531)" /gene="SLC52A2" CDS 3658..3723 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3767..3978 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3991..4259 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4288..4297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4318..4980 /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 5050..5103 /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 5104..5703 /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 5704..6292 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6366..6599 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6671..6792 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6832..6967 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7000..7016) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7024..7040 /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(7048..7078) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7093..7114 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7402..7990) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8161..9021) /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(9022..9126) /gene="bla" /label="AmpR promoter" primer_bind 9600..9616 /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 cagcacccac gcccgcccgt 2341 ccggtgctga cccacctgct ggtggctctc ttcggcatgg gctcctgggc tgcggtcaat 2401 gggatctggg tggagctacc tgtggtggtc aaagagcttc cagagggttg gagcctcccc 2461 tcttacgtct ctgtgcttgt ggctctgggg aacctgggtc tgctggtggt gaccctctgg 2521 aggaggctgg ccccaggaaa ggacgagcag gtccccatcc gggtggtgca ggtgctgggc 2581 atggtgggca cagccctgct ggcctctctg tggcaccatg tggccccagt ggcaggacag 2641 ttgcattctg tggccttctt agcactggcc tttgtgctgg cactggcatg ctgtgcctcg 2701 aatgtcactt tcctgccctt cttgagccac ctgccacctc gcttcttacg gtcattcttc 2761 ctgggtcaag gcctgagtgc cctgctgccc tgcgtgctgg ccctagtgca gggtgtgggc 2821 cgcctcgagt gcccgccagc ccccatcaac ggcacccctg gccccccgct cgacttcctt 2881 gagcgttttc ccgccagcac cttcttctgg gcactgactg cccttctggt cgcttcagct 2941 gctgccttcc agggtcttct gctgctgttg ccgccaccac catctgtacc cacaggggag 3001 ttaggatcag gcctccaggt gggagcccca ggagcagagg aagaggtgga agagtcctca 3061 ccactgcaag agccaccaag ccaggcagca ggcaccaccc ctggtccaga ccctaaggcc 3121 tatcagcttc tatcagcccg cagtgcctgc ctgctgggcc tgttggccgc caccaacgcg 3181 ctgaccaatg gcgtgctgcc tgccgtgcag agcttttcct gcttacccta cgggcgtctg 3241 gcctaccacc tggctgtggt gctgggcagt gctgccaatc ccctggcctg cttcctggcc 3301 atgggtgtgc tgtgcaggtc cttggcaggg ctgggcggcc tctctctgct gggcgtgttc 3361 tgtgggggct acctgatggc gctggcagtc ctgagcccct gcccgcccct ggtgggcacc 3421 tcggcggggg tggtcctcgt ggtgctgtcg tgggtgctgt gtcttggcgt gttctcctac 3481 gtgaaggtgg cagccagctc cctgctgcat ggcgggggcc ggccggcatt gctggcagcc 3541 ggcgtggcca tccaggtggg ctctctgctc ggcgctgttg ctatgttccc cccgaccagc 3601 atctatcacg tgttccacag cagaaaggac tgtgcagacc cctgtgactc cggatccgac 3661 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 3721 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3781 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 3841 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 3901 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 3961 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 4021 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 4081 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4141 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4201 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4261 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4321 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4381 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4441 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4501 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4561 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4621 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4681 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4741 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4801 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 4861 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 4921 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 4981 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 5041 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 5101 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5161 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5221 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5281 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5341 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5401 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5461 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5521 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5581 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5641 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5701 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5761 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5821 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5881 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5941 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 6001 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 6061 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 6121 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6181 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6241 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 6301 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 6361 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg 6421 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6481 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6541 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6601 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 6661 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6721 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6781 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc 6841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 6901 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 6961 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg 7021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 7081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7201 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7801 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7921 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7981 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 8041 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 8101 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8161 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8221 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8281 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8341 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8401 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8461 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8521 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8581 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8641 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8701 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8761 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8821 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8881 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8941 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 9001 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 9061 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9121 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 9181 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9241 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9301 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9361 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 9421 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 9481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 9541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 9601 taaaacgacg gccagtgcca agctg //