LOCUS V040961 9493 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040961 VERSION V040961 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..3519 /label="CRTAP(NM_006371)" /note="CRTAP(NM_006371)" /gene="CRTAP" CDS 3526..3591 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3635..3846 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3859..4127 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4156..4165 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4186..4848 /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 4918..4971 /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 4972..5571 /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 5572..6160 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6234..6467 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6539..6660 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6700..6835 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6868..6884) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6892..6908 /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(6916..6946) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6961..6982 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7270..7858) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8029..8889) /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(8890..8994) /gene="bla" /label="AmpR promoter" primer_bind 9468..9484 /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 agccggggcg ccggggggcc 2341 gcggcgctgc tagcgctgct gtgcgtggcc tgcgcgctgc gcgccgggcg cgcccaatac 2401 gaacgctaca gcttccgcag cttcccacgg gacgagctga tgccgctcga gtcggcctac 2461 cggcacgcgc tggacaagta cagcggcgag cactgggccg agagcgtggg ctacctggag 2521 atcagcctgc ggctgcaccg cttgctgcgc gacagcgagg ccttctgcca ccgcaactgc 2581 agcgccgcgc cgcagcccga gcccgccgcc ggcctcgcca gctatcccga gctgcgcctc 2641 ttcgggggcc tgctgcgccg cgcgcactgc ctcaagcgct gcaagcaggg cctgccagcc 2701 ttccgccagt cccagcccag ccgcgaggtg ctggcggact tccagcgccg cgagccctac 2761 aagttcctgc agttcgctta cttcaaggca aataatctcc ccaaagccat cgccgctgct 2821 cacacctttc tactgaagca tcctgatgac gaaatgatga agaggaacat ggcatattat 2881 aagagcctgc ctggtgccga ggactacatt aaagacctgg aaaccaagtc atatgaaagc 2941 ctgttcatcc gagcagtgcg ggcatacaac ggtgagaact ggagaacatc catcacagac 3001 atggagctgg cccttcccga cttcttcaaa gccttttacg agtgtctcgc agcctgcgag 3061 ggttccaggg agatcaagga cttcaaggat ttctaccttt ccatagcaga tcattatgta 3121 gaagttctgg aatgcaaaat acagtgtgaa gagaacctca ccccagttat aggaggctat 3181 ccggttgaga aatttgtggc taccatgtat cattacttgc agtttgccta ttataagttg 3241 aacgacctga agaatgcagc cccctgtgca gtcagctatc tgctctttga tcagaatgac 3301 aaggtcatgc agcagaacct ggtgtattac cagtaccaca gggacacttg gggcctctcg 3361 gatgagcact tccagcccag acctgaagca gttcagttct ttaatgtgac cacactccag 3421 aaggagctgt atgactttgc taaggaaaat ataatggatg atgatgaggg agaagttgtg 3481 gaatatgtgg atgacctctt ggaactggag gagaccagcg gatccgacta caaagaccat 3541 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3601 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3661 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3721 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3781 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3841 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3901 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3961 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4021 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4081 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4141 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4201 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4261 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4321 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4381 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4441 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4501 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4561 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4621 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4681 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4741 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4801 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4861 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4921 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4981 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5041 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5101 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5161 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5221 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5281 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5341 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5401 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5461 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5521 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5581 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5641 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5701 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5761 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5821 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5881 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5941 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6001 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6061 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6121 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6181 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6241 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6301 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6361 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6421 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6481 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6541 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6601 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6661 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6721 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6781 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6841 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6901 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6961 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7021 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7081 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7141 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7681 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7921 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7981 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8041 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8101 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8161 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8221 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8281 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8341 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8401 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8461 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8521 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8581 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8641 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8701 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8761 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8821 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8881 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8941 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9001 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9061 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9121 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9181 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9241 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9301 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9361 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9421 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9481 cagtgccaag ctg //