LOCUS V039975 8530 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039975 VERSION V039975 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3384 /label="HOMER3(NM_001145722)" /note="HOMER3(NM_001145722)" /gene="HOMER3" 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)" CDS 4019..4618 /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 4625..5213 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5287..5520 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5592..5713 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5732..5867 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5905..5921) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5929..5945 /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(5953..5983) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5998..6019 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6307..6895) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7066..7926) /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(7927..8031) /gene="bla" /label="AmpR promoter" primer_bind 8505..8521 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgtccaca gccagggagc agccaatctt cagcacacgg 2341 gcgcacgtgt tccaaattga cccagccacc aagcgaaact ggatcccagc gggcaagcac 2401 gcactcactg tctcctattt ctacgatgcc acccgcaatg tgtaccgcat catcagcatc 2461 ggaggcgcca aggccatcat caacagcact gtcactccca acatgacctt caccaaaact 2521 tcccagaagt tcgggcagtg ggccgacagt cgcgccaaca cagtctacgg cctgggcttt 2581 gcctctgaac agcatctgac acagtttgcc gagaagttcc aggaagtgaa ggaagcagcc 2641 aggctggcca gggagaaatc tcaggatggc ggggagctca ccagtccagc cctggggctc 2701 gcctcccacc aggtgccccc gagccctctc gtcagtgcca acggccccgg cgaggaaaaa 2761 ctgttccgca gccagagcgc tgatgccccc ggccccacag agcgcgagcg gctaaagaag 2821 atgttgtctg agggctccgt gggcgaggta cagtgggagg ccgagttttt cgcactgcag 2881 gacagcaaca acaagctggc aggcgccctg cgagaggcca acgccgccgc agcccagtgg 2941 aggcagcagc tggaggctca gcgtgcagag gccgagcggc tgcggcagcg ggtggctgag 3001 ctggaggctc aggcagcttc agaggtgacc cccaccggtg agaaggaggg gctgggccag 3061 ggccagtcgc tggaacagct ggaagctctg gtgcaaacca aggaccagga gattcagacc 3121 ctgaagagtc agactggggg gccccgcgag gccctggagg ctgccgagcg tgaggagact 3181 cagcagaagg tgcaggacct ggagacccgc aatgcggagt tggagcacca gctgcgggcg 3241 atggagcgca gcctggagga ggcacgggca gagcgggagc gggcgcgggc tgaggtgggc 3301 cgggcagcgc agctgctgga cgtcagcctg tttgagctga gtgagctgcg tgagggcctg 3361 gcccgcctgg ctgaggctgc gcccggatcc 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 tacgctagat 4021 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 4081 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 4141 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4201 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4261 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4321 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4381 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4441 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4501 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4561 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 4621 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4681 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4741 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4801 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 4861 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 4921 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 4981 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5041 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5101 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5161 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5221 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5281 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5341 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5401 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5461 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5521 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5581 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5641 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5701 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 5761 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 5821 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 5881 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 5941 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6001 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6061 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6121 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6181 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6241 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6301 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6361 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6421 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6481 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6541 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6601 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6661 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6721 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6781 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 6841 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 6901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 6961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 7861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 7921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 7981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8041 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8101 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8161 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8221 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8281 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8341 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8401 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8461 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8521 tgccaagctg //