LOCUS V047048 9578 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047048 VERSION V047048 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3741 /codon_start=1 /gene="AKT1" /product="human serine/threonine protein kinase activated by growth factors" /protein_id="" /transl_table=1 /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM ECVDSERRPHFPQFSYSASGTA" /label="Akt1" CDS 3781..4500 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced CFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG IKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="ECFP" promoter 4548..4759 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4772..5040 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5067..5666 /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 5673..6261 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6335..6568 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6640..6761 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6780..6915 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6953..6969) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6977..6993 /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(7001..7031) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7046..7067 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7355..7943) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8114..8974) /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(8975..9079) /gene="bla" /label="AmpR promoter" primer_bind 9553..9569 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgagcgac gtggctattg tgaaggaggg ttggctgcac 2341 aaacgagggg agtacatcaa gacctggcgg ccacgctact tcctcctcaa gaatgatggc 2401 accttcattg gctacaagga gcggccgcag gatgtggacc aacgtgaggc tcccctcaac 2461 aacttctctg tggcgcagtg ccagctgatg aagacggagc ggccccggcc caacaccttc 2521 atcatccgct gcctgcagtg gaccactgtc atcgaacgca ccttccatgt ggagactcct 2581 gaggagcggg aggagtggac aaccgccatc cagactgtgg ctgacggcct caagaagcag 2641 gaggaggagg agatggactt ccggtcgggc tcacccagtg acaactcagg ggctgaagag 2701 atggaggtgt ccctggccaa gcccaagcac cgcgtgacca tgaacgagtt tgagtacctg 2761 aagctgctgg gcaagggcac tttcggcaag gtgatcctgg tgaaggagaa ggccacaggc 2821 cgctactacg ccatgaagat cctcaagaag gaagtcatcg tggccaagga cgaggtggcc 2881 cacacactca ccgagaaccg cgtcctgcag aactccaggc accccttcct cacagccctg 2941 aagtactctt tccagaccca cgaccgcctc tgctttgtca tggagtacgc caacgggggc 3001 gagctgttct tccacctgtc ccgggagcgt gtgttctccg aggaccgggc ccgcttctat 3061 ggcgctgaga ttgtgtcagc cctggactac ctgcactcgg agaagaacgt ggtgtaccgg 3121 gacctcaagc tggagaacct catgctggac aaggacgggc acattaagat cacagacttc 3181 gggctgtgca aggaggggat caaggacggt gccaccatga agaccttttg cggcacacct 3241 gagtacctgg cccccgaggt gctggaggac aatgactacg gccgtgcagt ggactggtgg 3301 gggctgggcg tggtcatgta cgagatgatg tgcggtcgcc tgcccttcta caaccaggac 3361 catgagaagc tttttgagct catcctcatg gaggagatcc gcttcccgcg cacgcttggt 3421 cccgaggcca agtccttgct ttcagggctg ctcaagaagg accccaagca gaggcttggc 3481 gggggctccg aggacgccaa ggagatcatg cagcatcgct tctttgccgg tatcgtgtgg 3541 cagcacgtgt acgagaagaa gctcagccca cccttcaagc cccaggtcac gtcggagact 3601 gacaccaggt attttgatga ggagttcacg gcccagatga tcaccatcac accacctgac 3661 caagatgaca gcatggagtg tgtggacagc gagcgcaggc cccacttccc ccagttctcc 3721 tactcggcca gcggcacggc cctctcgagt ggcggtggag gaagcggcgg aggtggatct 3781 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 3841 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 3901 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 3961 ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc gctaccccga ccacatgaag 4021 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 4081 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 4141 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 4201 aagctggagt acaactacat cagccacaac gtctatatca ccgccgacaa gcagaagaac 4261 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 4321 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 4381 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 4441 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga 4501 ggatccgcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac 4561 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag 4621 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 4681 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 4741 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg 4801 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg 4861 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg 4921 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct 4981 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc 5041 caagctgtga ccggcgccta cgctagatga ccgagtacaa gcccacggtg cgcctcgcca 5101 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 5161 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 5221 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 5281 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 5341 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 5401 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 5461 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 5521 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 5581 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 5641 gcatgacccg caagcccggt gcctgagtcg acaatcaacc tctggattac aaaatttgtg 5701 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 5761 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 5821 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 5881 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 5941 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 6001 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 6061 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 6121 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 6181 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 6241 ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta 6301 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga 6361 aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg 6421 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 6481 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 6541 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 6601 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat 6661 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 6721 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta 6781 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 6841 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 6901 gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc 6961 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7021 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7081 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7141 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7201 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7261 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7321 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7381 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7441 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7501 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 7801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 7861 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 7921 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 7981 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8041 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8101 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8161 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8221 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8281 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8341 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8401 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8461 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 8521 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 8581 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 8641 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 8701 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 8761 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 8821 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 8881 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 8941 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9001 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9061 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca 9121 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc 9181 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt 9241 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg 9301 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata 9361 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc 9421 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 9481 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc 9541 agtcacgacg ttgtaaaacg acggccagtg ccaagctg //