LOCUS V030390 9364 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030390 VERSION V030390 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 2303..3478 /label="FEZ1(NM_005103)" /note="FEZ1(NM_005103)" /gene="FEZ1" promoter 3523..3734 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3747..4015 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4044..4053 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4074..4736 /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 4806..4859 /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 4860..5459 /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 5460..6047 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6121..6354 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6426..6547 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6566..6701 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6739..6755) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6763..6779 /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(6787..6817) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6832..6853 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7141..7729) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7900..8760) /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(8761..8865) /gene="bla" /label="AmpR promoter" primer_bind 9339..9355 /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 gaattcgcca ccatggaggc cccactggtg agtctggatg aagagtttga 2341 ggaccttcga ccctcctgct cggaggaccc ggaggagaag ccccagtgtt tctatggttc 2401 atctccccac catctcgagg acccctccct ctccgagctt gagaattttt cttccgaaat 2461 aatcagcttc aagtccatgg aggacctcgt aaatgaattt gatgagaagc tcaatgtctg 2521 ctttcggaac tacaacgcca agaccgagaa cctagctccc gtgaagaacc agttacagat 2581 ccaagaggag gaggagaccc ttcaggacga ggaggtttgg gatgctctga cagacaatta 2641 catcccttca ctctcagaag actggaggga tccaaacatc gaggctctga atggcaactg 2701 ctctgacact gagatccatg agaaagaaga ggaagagttc aatgagaaga gtgaaaatga 2761 ttccggtatc aacgaggagc ctctgctcac agcagatcag gtaattgagg agattgagga 2821 aatgatgcag aactccccag accctgagga agaagaggag gttctggaag aagaggatgg 2881 aggagaaact tcctcccagg cagactcggt cctcctgcag gagatgcagg cattgacaca 2941 gaccttcaac aacaactggt cctatgaagg gctgaggcac atgtctgggt ctgagctgac 3001 cgagctgctg gaccaggtgg agggtgccat ccgtgacttc tcggaggagc tggtgcagca 3061 gctggcccgc cgggacgagc tggagtttga gaaggaagtg aagaactcct ttatcacggt 3121 gcttattgag gttcagaaca agcagaagga gcagcgagaa ctgatgaaaa agaggcggaa 3181 agagaaaggg ctgagcctgc agagcagccg gatagagaag ggaaaccaga tgcctctcaa 3241 gcgcttcagc atggaaggca tctccaacat tctgcagagt ggcatccgcc agacctttgg 3301 ctcctcagga actgacaaac agtatctgaa cacagtcatt ccttacgaga agaaagcctc 3361 tcctccctca gtggaagacc tgcagatgct gacaaacatt ctctttgcca tgaaggagga 3421 taatgagaag gtgcctactt tgctaacgga ctacatttta aaagtgctct gccctaccta 3481 agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc 3541 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt 3601 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg 3661 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg 3721 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc 3781 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg 3841 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt 3901 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc 3961 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc 4021 tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca 4081 tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg 4141 gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg 4201 ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg 4261 gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca 4321 ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc 4381 gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg 4441 acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc 4501 ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg 4561 gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca 4621 tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca 4681 ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat 4741 cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat 4801 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta 4861 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac 4921 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc 4981 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca 5041 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga 5101 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt 5161 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc 5221 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca 5281 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg 5341 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg 5401 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa 5461 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5521 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5581 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5641 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 5701 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 5761 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5821 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 5881 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5941 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc 6001 gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa 6061 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6121 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 6181 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6241 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6301 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6361 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6421 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6541 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6721 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6781 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6841 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6901 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7561 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8881 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8941 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9001 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9061 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9121 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9181 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9241 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9301 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9361 gctg //