LOCUS V024966 9788 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024966 VERSION V024966 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..3831 /label="SLC37A2(NM_198277)" /note="SLC37A2(NM_198277)" /gene="SLC37A2" CDS 3838..3903 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3947..4158 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4171..4439 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4468..4477 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4498..5160 /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 5230..5283 /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 5284..5883 /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 5884..6471 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6545..6778 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6850..6971 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6990..7125 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7163..7179) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7187..7203 /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(7211..7241) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7256..7277 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7565..8153) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8324..9184) /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(9185..9289) /gene="bla" /label="AmpR promoter" primer_bind 9763..9779 /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 ggtcctccct ggctccggga 2341 gtctggttct tccgggcctt ctccagggac agctggttcc gaggcctcat cctgctgctg 2401 accttcctaa tttacgcctg ctatcacatg tccaggaagc ctatcagtat cgtcaagagc 2461 cgtctgcacc agaactgctc ggagcagatc aaacccatca atgatactca cagtctcaat 2521 gacaccatgt ggtgcagctg ggccccattt gacaaggaca actataagga gttactaggg 2581 ggcgtggaca acgccttcct catcgcctat gccatcggca tgttcatcag tggggttttt 2641 ggggagcggc ttccgctccg ttactacctc tcagctggaa tgctgctcag tggccttttc 2701 acctcgctct ttggcctggg atatttctgg aacatccacg agctctggta ctttgtggtc 2761 atccaggtct gtaatggact cgtccagacc acaggctggc cctctgtggt gacctgtgtt 2821 ggcaactggt tcgggaaggg gaagcggggg ttcatcatgg gcatctggaa ttcccacaca 2881 tctgtgggca acatcctggg ctccctgatc gccggcatct gggtgaacgg gcagtggggc 2941 ctgtcgttca tcgtgcctgg catcattact gccgtcatgg gcgtcatcac cttcctcttc 3001 ctcatcgaac acccagaaga tgtggactgc gcccctcctc agcaccacgg tgagccagct 3061 gagaaccagg acaaccctga ggaccctggg aacagtccct gctctatcag ggagagcggc 3121 cttgagactg tggccaaatg ctccaagggg ccatgcgaag agcctgctgc catcagcttc 3181 tttggggcgc tccggatccc aggcgtggtc gagttctctc tgtgtctgct gtttgccaag 3241 ctggtcagtt acaccttcct ctactggctg cccctctaca tcgccaatgt ggctcacttt 3301 agtgccaagg aggctgggga cctgtctaca ctcttcgatg ttggtggcat cataggcggc 3361 atcgtggcag ggctcgtctc tgactacacc aatggcaggg ccaccacttg ctgtgtcatg 3421 ctcatcttgg ctgcccccat gatgttcctg tacaactaca ttggccagga cgggattgcc 3481 agctccatag tgatgctgat catctgtggg ggcctggtca atggcccata cgcgctcatc 3541 accactgctg tctctgctga tctggggact cacaagagcc tgaagggcaa cgccaaagcc 3601 ctgtccacgg tcacggccat cattgacggc accggctcca taggtgcggc tctggggcct 3661 ctgctggctg ggctcatctc ccccacgggc tggaacaatg tcttctacat gctcatctct 3721 gccgacgtcc tagcctgctt gctcctttgc cggttagtat acaaagagat cttggcctgg 3781 aaggtgtccc tgagcagagg cagcggctct agtatggtcc taacccacca gggatccgac 3841 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 3901 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3961 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 4021 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 4081 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 4141 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 4201 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 4261 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4321 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4381 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4441 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4501 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4561 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4621 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4681 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4741 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4801 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4861 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4921 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4981 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 5041 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 5101 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 5161 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 5221 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 5281 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5341 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5401 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5461 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5521 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5581 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5641 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5701 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5761 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5821 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5881 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5941 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 6001 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 6061 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 6121 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 6181 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 6241 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 6301 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6361 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6421 ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct 6481 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6541 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6781 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6841 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6961 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 7021 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 7081 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 7141 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 7201 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7261 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7321 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7381 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7441 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7501 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7981 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 8041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 8101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 8161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 8221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 9001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 9061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 9121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 9181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9301 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9361 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9421 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9481 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9541 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9601 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9661 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9721 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9781 ccaagctg //