LOCUS V028976 8878 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028976 VERSION V028976 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..2904 /label="Cd300e(NM_172050)" /note="Cd300e(NM_172050)" /gene="Cd300e" CDS 2911..2976 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3020..3231 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3244..3512 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3541..3550 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3571..4233 /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 4303..4356 /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 4357..4956 /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 4957..5545 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5619..5852 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5924..6045 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6085..6220 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6253..6269) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6277..6293 /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(6301..6331) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6346..6367 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6655..7243) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7414..8274) /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(8275..8379) /gene="bla" /label="AmpR promoter" primer_bind 8853..8869 /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 gccaccatga ggctatgtgc aggtctgctc 2341 cttctctgct tccaaggttg tttgtctctg acgggccctg gctctgtgtc tggctacgta 2401 ggaggctctc tccgtgtgca gtgtcaatat agtccatcat ataagggcta tatgaaatac 2461 tggtgccgag gaccgcatga cacgacatgt aaaactattg tagaaaccga cggaagtgag 2521 aaagaaaaga ggagtggccc ggtgtccatc agagaccatg cttcgaactc caccatcaca 2581 gtgatcatgg aggacctcag cgaagacaat gctgggtctt actggtgcaa gattcagact 2641 tcctttatct gggattcgtg gtcacgtgat ccatcggtca gtgtaagggt gaatgttttt 2701 ccagccacaa ctcctacact cccagccaca actgctatcc tcccactagt gaattctggg 2761 cagaacctga ggattagtac taatgtgatg ttcatcttcc aactgtggtc cctgctcagc 2821 agcatccagt tccaggtcct ggtcttcctg aagctgcctc tgtttctgag catgctctgt 2881 gctatcttct gggtgaacag acttggatcc gactacaaag accatgacgg tgattataaa 2941 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3001 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3061 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3241 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3301 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3361 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3421 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3481 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3541 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 3601 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 3661 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 3721 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 3781 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 3841 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 3901 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 3961 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4021 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4081 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4141 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4201 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4261 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4321 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4381 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4441 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4501 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4561 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 4621 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 4681 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 4741 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 4801 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 4861 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 4921 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 4981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5101 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5161 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5221 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5281 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5341 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5401 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5461 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5521 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 5581 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 5641 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 5701 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 5761 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 5821 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 5881 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 5941 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6001 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6061 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6181 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6241 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6301 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6361 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6421 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6481 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6541 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6601 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6661 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6721 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6781 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6841 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6901 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6961 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7021 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7081 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7141 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7201 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7261 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7321 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7381 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7441 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7501 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7561 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7621 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7681 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7741 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7801 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7861 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7921 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7981 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8041 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8101 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8161 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8221 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8281 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8341 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8401 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8461 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8521 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8581 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8641 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8701 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8761 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8821 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //