LOCUS V029303 9964 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029303 VERSION V029303 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..3990 /label="Fancc(NM_007985)" /note="Fancc(NM_007985)" /gene="Fancc" CDS 3997..4062 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4106..4317 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4330..4598 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4627..4636 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4657..5319 /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 5389..5442 /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 5443..6042 /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 6043..6631 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6705..6938 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7010..7131 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7171..7306 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7339..7355) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7363..7379 /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(7387..7417) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7432..7453 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7741..8329) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8500..9360) /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(9361..9465) /gene="bla" /label="AmpR promoter" primer_bind 9939..9955 /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 ctcaggagtc tgcagacctt 2341 gcttctgact gtcagtcttg gctgcagaag ctttctgcat gggaacaggc ctcttctgag 2401 gaaacccaga aggacacttg tcttcacttg tccgggttcc aggagttcct gaggcagatg 2461 tatgaaatct tgaaggagat ggattctgat gcaatcctgg aaaggttccc cacaattggt 2521 caactgttgg caaaagcttg ttggaatcct ctcatcttag catatgatga aagccaaaaa 2581 attgtaatat ggtgcttatg ttgtctgatg aacaaagaac ctcggacttc tgcagagtca 2641 ggacttaact cgtggatccg gggtttgtta tctcatgtac tttctgcatt cagattcgac 2701 atgaaagaag tttgtctttt taccaaaagt cttggatatg agtctattga ttactatcct 2761 agtttgctta aaaatatggt tttgtcatta gtgtctgagc tcagagagag tcatcttaat 2821 ggactgagca ctcaaagtcg gatggctcct gagcgcatga tgtccctgtc agaagtttgt 2881 gtccctcttg tcactctgcc tgatatggaa cccctggtag aggctctact cacctaccat 2941 ggacatgagc cccaggaagt cctggctcct gagttcttcg aagctgtaaa tgaggccttc 3001 ttgtcggaaa aaattgttct acccacgtcc tctgtggtca gcctctggtt tcggcatctc 3061 cccagtcttg aaaaagcaac gctgcatctt tttgaaaagc ttttctccag caagataatt 3121 tgcctgagaa ggatggagtg ctgtataaga gagtcattcc tgcctcaagc agcctgccaa 3181 cctgccatct tcagaattgt tgatgaaatg ttcaggtttg tgctgctgga aactgacgga 3241 gccccagaag tactagctgc tcttcaggtt ttcacatcgt gcttggtaga agctctgaaa 3301 aaagaaaaca agcagctgac gtttgccctc aggacctact ttccttacgg tgctccatgt 3361 cttgctgcag cgctgtccca gcaccctgaa gcaatcccac agggacaccg gctccagcct 3421 ctgctgcaca tttcccaact cctcagagaa gcagttgaag actgtactcg tgggtctccg 3481 cgaaatccct ttgagagctg gtttttgttt gttcactttg gaggatgggt tgacctggct 3541 gtggcagagt tactgctgag ggaggaagct gagcctcctg ctggcctgct gtggctcttg 3601 gtgttctatt acagcccaca ggatgggagt cagcagagag agcagagcat ggtggagctg 3661 aaggtattaa tcaaccgtct cctgatgctg ctcagaagcg gccccctctc agctactgat 3721 ctgcaggaag cagctgagag tcccagtgga gaccccagac cacctgtatg tggacagctg 3781 gtcagacgcc ttcttcttag tctcttgctc tggaccccag aaggccatgc aattgtctgg 3841 gaagctgtca cccatatggc ccacacggat gctgtaatcc atgagattat tggttttctt 3901 gaccagacct tgtacagatc acaacatctt tgtgttgaag cctcgagaaa actggccaga 3961 gacctcctaa aggagctgca agcccaggtc ggatccgact acaaagacca tgacggtgat 4021 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 4081 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 4141 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 4201 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 4261 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4321 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4381 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4441 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4501 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4561 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4621 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4681 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4741 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4801 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4861 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4921 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4981 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 5041 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 5101 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 5161 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 5221 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 5281 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 5341 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 5401 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5461 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5521 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5581 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5641 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5701 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5761 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5821 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5881 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5941 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 6001 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 6061 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 6121 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 6181 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6241 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6301 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6361 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6421 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6481 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6541 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6601 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6661 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6721 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6781 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6841 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6901 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6961 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 7021 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 7081 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 7141 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 7201 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 7261 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 7321 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 8041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 8101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8161 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 9001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 9061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9481 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9541 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9601 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9661 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9721 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9781 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9841 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9901 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9961 gctg //