LOCUS V042929 8765 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042929 VERSION V042929 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..2859 /label="LEMD1(NM_001199050)" /note="LEMD1(NM_001199050)" /gene="LEMD1" promoter 2924..3135 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3148..3416 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3445..3454 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3475..4137 /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 4207..4260 /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 4261..4860 /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 4861..5448 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5522..5755 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5827..5948 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5967..6102 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6140..6156) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6164..6180 /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(6188..6218) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6233..6254 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6542..7130) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7301..8161) /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(8162..8266) /gene="bla" /label="AmpR promoter" primer_bind 8740..8756 /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 tggatgtgaa gtgtctgagt 2341 gactgtaaat tgcagaacca acttgagaag cttggatttt cacctggccc aatactacct 2401 tccaccagaa agttgtatga aaaaaagtta gtacagttgt tggtctcacc tccctgtgca 2461 ccacctgtga tgaatggacc cagagagctg gatggagcgc aggacagtga tgacagcgaa 2521 gagcttaata tcattttgca aggaaatatc atactctcaa cagaaaaaag caagaaactc 2581 aaaaaatggc ctgaggcttc caccactaaa cgcaaagctg tagataccta ttgcttggat 2641 tataagcctt ccaagggaag aaggtgggct gcaagagcac caagcaccag aatcacatat 2701 gggactatca ccaaagagag agactactgc gcggaagacc agactatcga gagctggaga 2761 gaagaaggtt tcccagtggg cttgaagctt gctgtgcttg gtattttcat cattgtggtg 2821 tttgtctacc tgactgtgga aaataagtcg ctgtttggtt aaggatccct gcgatatcca 2881 gcgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 2941 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3001 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3061 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3121 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 3181 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 3241 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 3301 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 3361 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 3421 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 3481 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 3541 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 3601 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 3661 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 3721 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 3781 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 3841 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 3901 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 3961 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4021 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4081 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4141 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 4201 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 4261 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 4321 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 4381 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 4441 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 4501 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 4561 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 4621 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 4681 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 4741 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 4801 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 4861 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 4921 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 4981 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5041 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5101 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5161 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5221 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 5281 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 5341 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 5401 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 5461 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5521 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 5581 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5641 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 5701 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 5761 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 5821 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 5881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 5941 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6001 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6061 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6121 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6181 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6241 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6301 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6361 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6421 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6481 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6541 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6601 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6661 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6721 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 6781 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 6841 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 6901 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 6961 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7021 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7081 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7141 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7201 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7261 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7321 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7381 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7441 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7501 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7561 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7621 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7681 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 7741 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 7801 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 7861 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 7921 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 7981 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8041 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8101 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8161 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8221 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8281 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8341 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 8401 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 8461 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 8521 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8581 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 8641 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 8701 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 8761 agctg //