LOCUS V041416 8938 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041416 VERSION V041416 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..2964 /label="Ppib(NM_011149)" /note="Ppib(NM_011149)" /gene="Ppib" CDS 2971..3036 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3080..3291 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3304..3572 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3601..3610 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3631..4293 /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 4363..4416 /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 4417..5016 /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 5017..5605 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5679..5912 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5984..6105 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6145..6280 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6313..6329) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6337..6353 /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(6361..6391) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6406..6427 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6715..7303) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7474..8334) /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(8335..8439) /gene="bla" /label="AmpR promoter" primer_bind 8913..8929 /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 tgcgcctctc ggagcgcaat 2341 atgaaggtgc tcttcgccgc cgccctcatc gtgggctccg tcgtcttcct tttgctgccc 2401 ggaccctccg tggccaacga taagaagaag ggacctaaag tcacagtcaa ggtatacttt 2461 gatttacaaa ttggagatga atctgtagga cgagtcgtct ttggactctt tggaaagact 2521 gttccaaaaa cagtggataa ttttgtagcc ttagctacag gagagaaagg atttggctac 2581 aaaaacagca agttccatcg tgtcatcaag gacttcatga tccagggtgg agacttcacc 2641 aggggagatg gcacaggagg aaagagcatc tatggtgagc gcttcccaga tgagaacttc 2701 aagctgaagc actacgggcc tggctgggtg agcatggcca atgcaggcaa agacaccaat 2761 ggctcacagt tcttcataac cacagtcaag acctcctggc tggatggcaa gcatgtggtt 2821 ttcggcaaag ttctagaggg catggatgtg gtacggaagg tggagagcac caagacagac 2881 agccgggaca agccactgaa ggatgtcatc attgtcgact ccggcaagat cgaagtggag 2941 aaacccttcg ccattgccaa ggagggatcc gactacaaag accatgacgg tgattataaa 3001 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3061 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3121 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3181 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3241 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3301 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3361 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3421 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3481 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3541 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3601 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 3661 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 3721 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 3781 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 3841 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 3901 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 3961 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4021 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4081 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4141 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4201 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4261 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4321 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4381 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4441 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4501 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4561 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4621 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 4681 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 4741 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 4801 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 4861 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 4921 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 4981 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5041 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5101 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5161 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5221 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5281 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5341 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5401 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5461 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5521 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5581 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 5641 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 5701 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 5761 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 5821 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 5881 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 5941 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6001 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6061 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6121 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6181 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6241 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6301 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6361 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6481 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6541 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6601 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6661 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6721 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6901 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7141 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7261 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7321 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7381 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7441 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7501 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7561 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7621 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7681 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7741 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7801 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7861 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7921 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7981 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8041 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8101 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8161 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8221 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8281 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8341 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8461 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8521 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8581 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8641 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8701 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8761 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8821 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8881 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //