LOCUS V032092 8522 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032092 VERSION V032092 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 1960..2171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2184..2452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 2498..2725 /label="Cd24a(NM_009846)" /note="Cd24a(NM_009846)" /gene="Cd24a" CDS 2762..3481 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" promoter 3491..3990 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4011..4610 /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 4617..5205 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5279..5512 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5584..5705 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5724..5859 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5897..5913) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5921..5937 /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(5945..5975) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5990..6011 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6299..6887) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7058..7918) /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(7919..8023) /gene="bla" /label="AmpR promoter" primer_bind 8497..8513 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca 1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc 2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta 2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct 2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc 2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc 2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt 2461 gaccggcgcc tactctagag ctagcgaatt cgccaccatg ggcagagcga tggtggccag 2521 gctagggctg gggttgctgc ttctggcact gctcctaccc acgcagattt actgcaacca 2581 aacatctgtt gcaccgtttc ccggtaacca gaatatttct gcttccccaa atccaagtaa 2641 cgctaccacc agagggggtg gcagctccct gcagtccaca gctggtctcc tggctctctc 2701 tctctctctt ctacatctct actgtggatc cggcggtgga ggaagcggcg gaggtggatc 2761 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 2821 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 2881 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 2941 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa 3001 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 3061 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 3121 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 3181 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 3241 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 3301 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 3361 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 3421 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta 3481 ggcggccgct gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca 3541 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc 3601 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct 3661 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg 3721 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg 3781 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg 3841 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc 3901 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg 3961 ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt 4021 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg 4081 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg 4141 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 4201 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 4261 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 4321 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 4381 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 4441 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 4501 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 4561 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc 4621 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 4681 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 4741 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 4801 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 4861 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 4921 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 4981 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct 5041 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 5101 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 5161 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga 5221 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 5281 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 5341 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 5401 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 5461 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 5521 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 5581 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 5641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5701 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 5761 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 5821 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 5881 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 5941 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6001 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 6061 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 6121 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 6181 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 6241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 6301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 6361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 6421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 6481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 6541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 6601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 6661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 6721 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 6781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 6841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 6901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 6961 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 7081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 7141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 7201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 7261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 7321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 7381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 7441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 7501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 7561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 7621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 7681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 7741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 7801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 7861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 7921 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 7981 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 8041 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 8101 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 8161 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 8221 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 8281 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 8341 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 8401 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 8461 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 8521 tg //