LOCUS V035993 8368 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035993 VERSION V035993 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 1960..2171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2184..2452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 2495..2521 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2522..3775 /label="RAD52(NM_134424)" /note="RAD52(NM_134424)" /gene="RAD52" CDS 3791..3844 /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 3851..4450 /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 4463..5051 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5125..5358 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5430..5551 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5570..5705 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5743..5759) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5767..5783 /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(5791..5821) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5836..5857 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6145..6733) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6904..7764) /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(7765..7869) /gene="bla" /label="AmpR promoter" primer_bind 8343..8359 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca 1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc 2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta 2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct 2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc 2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc 2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt 2461 gaccggcgcc tactctagag ctagcgccac catgtaccca tacgacgtac ctgattatgc 2521 aatgtctggg actgaggaag caattcttgg aggacgtgac agccatcctg ctgctggcgg 2581 cggctcagtg ttatgctttg gacagtgcca gtacacagca gaagagtacc aggccatcca 2641 gaaggccctg aggcagaggc tgggcccaga atacataagt agccgcatgg ctggcggagg 2701 ccagaaggtg tgctacattg agggtcatcg ggtaattaat ctggccaatg agatgtttgg 2761 ttacaatggc tgggcacact ccatcacgca gcagaatgtg gattttgttg acctcaacaa 2821 tggcaagttc tacgtgggag tctgtgcatt tgtgagggtc cagctgaagg atggttcata 2881 tcatgaagat gttggttatg gtgttagtga gggcctcaag tccaaggctt tatctttgga 2941 gaaggcaagg aaggaggcgg tgacagacgg gctgaagcga gccctcagga gttttgggaa 3001 tgcacttgga aactgtattc tggacaaaga ctacctgaga tcactaaata agcttccacg 3061 ccagttgcct cttgaagtgg atttaactaa agcgaagaga caagatcttg aaccgtctgt 3121 ggaggaggca agatacaaca gctgccgacc gaacatggcc ctgggacacc cacagctgca 3181 gcaggtgacc tccccttcca gacccagcca tgctgtgata ccggcggacc aggactgcag 3241 ctcccgaagc ctgagctcat ccgccgtgga gagcgaggcc acgcaccagc ggaagctccg 3301 gcagaagcag ctgcagcagc agttccggga gcggatggag aagcagcagg ttcgagtctc 3361 cacgccgtca gctgagaaga gtgaggcagc gcctccggcc cctcctgtga cgcacagcac 3421 tcctgtaact gtctcagaac cactcctgga gaaagacttc cttgcaggag tgactcaaga 3481 attaatcaag actcttgaag acaactctga aaagtgggct gtgactcccg atgcagggga 3541 tggtgtggtc aagccctcgt ctagagcaga cccagcccag acctctgaca cattagcctt 3601 gaacaaccag atggtgaccc agaacaggac tccacacagc gtttgccacc agaaaccaca 3661 agcaaaatct ggatcttggg acctccaaac ttatagcgct gaccaacgca caacaggaaa 3721 ctgggaatct cataggaaga gccaggacat gaagaaaagg aaatatgatc catctggatc 3781 cgcggccgct gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg 3841 cccttccggg atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc 3901 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt 3961 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt 4021 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac 4081 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga 4141 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg 4201 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa 4261 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc 4321 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac 4381 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc 4441 cggtgcctga atctaggtcg acaatcaacc tctggattac aaaatttgtg aaagattgac 4501 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 4561 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 4621 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 4681 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 4741 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 4801 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 4861 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 4921 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 4981 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 5041 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 5101 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc 5161 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg 5221 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag 5281 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag 5341 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg 5401 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca 5461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 5521 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct 5581 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 5641 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 5701 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt 5761 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5821 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 5881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5941 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 6001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 6061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat 6601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 6721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 6781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 6841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 6901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 6961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 7021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 7081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 7141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 7201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 7261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 7321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 7381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 7441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 7501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 7621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 7681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 7741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 7801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 7861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat 7921 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga 7981 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc 8041 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg 8101 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga 8161 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct 8221 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 8281 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 8341 ttgtaaaacg acggccagtg ccaagctg //