LOCUS V023297 8944 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023297 VERSION V023297 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" regulatory 2297..2306 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2306..2329 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2330..3058 /label="CTHRC1(NM_138455)" /note="CTHRC1(NM_138455)" /gene="CTHRC1" promoter 3103..3314 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3327..3595 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3624..3633 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3654..4316 /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 4386..4439 /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 4440..5039 /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 5040..5627 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5701..5934 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6006..6127 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6146..6281 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6319..6335) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6343..6359 /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(6367..6397) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6412..6433 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6721..7309) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7480..8340) /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(8341..8445) /gene="bla" /label="AmpR promoter" primer_bind 8919..8935 /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 ccatggatta caaggatgac gacgataaga tgcgacccca 2341 gggccccgcc gcctccccgc agcggctccg cggcctcctg ctgctcctgc tgctgcagct 2401 gcccgcgccg tcgagcgcct ctgagatccc caaggggaag caaaaggcgc agctccggca 2461 gagggaggtg gtggacctgt ataatggaat gtgcttacaa gggccagcag gagtgcctgg 2521 tcgagacggg agccctgggg ccaatggcat tccgggtaca cctgggatcc caggtcggga 2581 tggattcaaa ggagaaaagg gggaatgtct gagggaaagc tttgaggagt cctggacacc 2641 caactacaag cagtgttcat ggagttcatt gaattatggc atagatcttg ggaaaattgc 2701 ggagtgtaca tttacaaaga tgcgttcaaa tagtgctcta agagttttgt tcagtggctc 2761 acttcggcta aaatgcagaa atgcatgctg tcagcgttgg tatttcacat tcaatggagc 2821 tgaatgttca ggacctcttc ccattgaagc tataatttat ttggaccaag gaagccctga 2881 aatgaattca acaattaata ttcatcgcac ttcttctgtg gaaggacttt gtgaaggaat 2941 tggtgctgga ttagtggatg ttgctatctg ggttggtact tgttcagatt acccaaaagg 3001 agatgcttct actggatgga attcagtttc tcgcatcatt attgaagaac taccaaaata 3061 agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc 3121 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt 3181 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg 3241 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg 3301 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc 3361 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg 3421 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt 3481 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc 3541 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc 3601 tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca 3661 tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg 3721 gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg 3781 ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg 3841 gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca 3901 ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc 3961 gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg 4021 acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc 4081 ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg 4141 gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca 4201 tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca 4261 ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat 4321 cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat 4381 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta 4441 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac 4501 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc 4561 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca 4621 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga 4681 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt 4741 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc 4801 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca 4861 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg 4921 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg 4981 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa 5041 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5101 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5161 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5221 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 5281 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 5341 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5401 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 5461 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5521 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc 5581 gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa 5641 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5701 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5761 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5821 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5881 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5941 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6001 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6061 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6121 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6301 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6361 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6421 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6481 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6541 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6601 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6661 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6721 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6781 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6841 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6901 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6961 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7021 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7081 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7141 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7201 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7261 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7321 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7381 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7441 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7501 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7561 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7621 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7681 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7741 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7801 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7861 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7921 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7981 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8041 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8101 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8161 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8221 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8281 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8341 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8401 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8461 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8521 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8581 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8641 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8701 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8761 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8821 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8881 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8941 gctg //