LOCUS V043299 9238 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043299 VERSION V043299 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..3264 /label="RDH12(NM_152443)" /note="RDH12(NM_152443)" /gene="RDH12" CDS 3271..3336 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3380..3591 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3604..3872 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3901..3910 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3931..4593 /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 4663..4716 /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 4717..5316 /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 5317..5905 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5979..6212 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6284..6405 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6445..6580 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6613..6629) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6637..6653 /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(6661..6691) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6706..6727 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7015..7603) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7774..8634) /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(8635..8739) /gene="bla" /label="AmpR promoter" primer_bind 9213..9229 /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 gccaccatgc tggtcacctt gggactgctc 2341 acctccttct tctcgttcct gtatatggta gctccatcca tcaggaagtt ctttgctggt 2401 ggagtgtgta gaacaaatgt gcagcttcct ggcaaggtag tggtgatcac tggcgccaac 2461 acgggcattg gcaaggagac ggccagagag ctcgctagcc gaggagcccg agtctatatt 2521 gcctgcagag atgtactgaa gggggagtct gctgccagtg aaatccgagt ggatacaaag 2581 aactcccagg tgctggtgcg gaaattggac ctatccgaca ccaaatctat ccgagccttt 2641 gctgagggct ttctggcaga ggaaaagcag ctccatattc tgatcaacaa tgcgggagta 2701 atgatgtgtc catattccaa gacagctgat ggctttgaaa cccacctggg agtcaaccac 2761 ctgggccact tcctcctcac ctacctgctc ctggagcggc taaaggtgtc tgcccctgca 2821 cgggtggtta atgtgtcctc ggtggctcac cacattggca agattccctt ccacgacctc 2881 cagagcgaga agcgctacag caggggtttt gcctattgcc acagcaagct ggccaatgtg 2941 ctttttactc gtgagctggc caagaggctc caaggcaccg gggtcaccac ctacgcagtg 3001 cacccaggcg tcgtccgctc tgagctggtc cggcactcct ccctgctctg cctgctctgg 3061 cggctcttct ccccctttgt caagacggca cgggaggggg cgcagaccag cctgcactgc 3121 gccctggctg agggcctgga gcccctgagt ggcaagtact tcagtgactg caagaggacc 3181 tgggtgtctc caagggcccg aaataacaaa acagctgagc gcctatggaa tgtcagctgt 3241 gagcttctag gaatccggtg ggagggatcc gactacaaag accatgacgg tgattataaa 3301 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3361 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3421 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3481 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3541 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3601 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3661 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3721 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3781 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3841 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3901 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 3961 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4021 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4081 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4141 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4201 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4261 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4321 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4381 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4441 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4501 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4561 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4621 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4681 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4741 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4801 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4861 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4921 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 4981 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5041 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5101 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5161 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5221 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5281 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5341 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5401 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5461 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5521 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5581 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5641 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5701 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5761 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5821 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5881 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 5941 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6001 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6061 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6121 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6181 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6241 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6301 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6361 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6421 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6481 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6541 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6601 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6661 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6721 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6781 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6841 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6901 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6961 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7021 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7081 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7141 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7201 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7261 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7321 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7381 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7441 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7501 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7561 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7621 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7681 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7741 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7801 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7861 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7921 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7981 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8041 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8101 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8161 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8221 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8281 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8341 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8401 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8461 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8521 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8581 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8641 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8701 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8761 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8821 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8881 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8941 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9001 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9061 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9121 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9181 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //