LOCUS V019660 8484 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019660 VERSION V019660 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 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..3013 /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" CDS 3029..3403 /codon_start=1 /gene="MAP1LC3B" /note="Atg8 family member found on autophagosomes" /product="human LC3 protein" /transl_table=1 /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD EDGFLYMVYASQETFGMKLSV" /label="LC3B" promoter 3454..3665 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3678..3946 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3973..4572 /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 4579..5167 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5241..5474 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5546..5667 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5686..5821 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5859..5875) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5883..5899 /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(5907..5937) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5952..5973 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6261..6849) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7020..7880) /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(7881..7985) /gene="bla" /label="AmpR promoter" primer_bind 8459..8475 /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 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 2341 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 2401 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 2461 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 2521 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 2581 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 2641 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 2701 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 2761 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 2821 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 2881 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 2941 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 3001 cgagctgtac aagggcggtg ggggaagcat gccgtcggag aagaccttca agcagcgccg 3061 caccttcgaa caaagagtag aagatgtccg acttattcga gagcagcatc caaccaaaat 3121 cccggtgata atagaacgat acaagggtga gaagcagctt cctgttctgg ataaaacaaa 3181 gttccttgta cctgaccatg tcaacatgag tgagctcatc aagataatta gaaggcgctt 3241 acagctcaat gctaatcagg ccttcttcct gttggtgaac ggacacagca tggtcagcgt 3301 ctccacacca atctcagagg tgtatgagag tgagaaagat gaagatggat tcctgtacat 3361 ggtctatgcc tcccaggaga cgttcgggat gaaattgtca gtgtaaggat ccgcggccgc 3421 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc 3481 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg 3541 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac 3601 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa 3661 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 3721 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 3781 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 3841 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 3901 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg 3961 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 4021 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 4081 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 4141 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 4201 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 4261 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 4321 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 4381 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 4441 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 4501 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 4561 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 4621 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 4681 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 4741 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 4801 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 4861 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 4921 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 4981 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 5041 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 5101 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 5161 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 5221 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 5281 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 5341 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 5401 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 5461 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 5521 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5641 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 5701 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 5761 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 5821 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 5881 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 5941 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 6001 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 6061 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6121 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6181 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6241 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6301 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6361 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 6421 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 6481 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 6541 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 6601 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 6661 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 6721 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 6781 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 6841 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 6901 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 6961 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 7021 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 7081 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 7141 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 7201 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 7261 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 7321 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 7381 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 7441 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 7501 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 7561 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 7621 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 7681 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 7741 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 7801 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 7861 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 7921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 7981 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 8041 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 8101 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 8161 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 8221 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 8281 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 8341 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 8401 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 8461 aaaacgacgg ccagtgccaa gctg //