LOCUS V029424 8648 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029424 VERSION V029424 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..3018 /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 3055..3573 /label="CRYAA(NM_000394)" /note="CRYAA(NM_000394)" /gene="CRYAA" promoter 3618..3829 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3842..4110 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4137..4736 /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 4743..5331 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5405..5638 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5710..5831 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5850..5985 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6023..6039) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6047..6063 /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(6071..6101) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6116..6137 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6425..7013) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7184..8044) /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(8045..8149) /gene="bla" /label="AmpR promoter" primer_bind 8623..8639 /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 catggtgagc aagggcgagg agctgttcac cggggtggtg 2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggac 3061 gtgaccatcc agcacccctg gttcaagcgc accctggggc ccttctaccc cagccggctg 3121 ttcgaccagt ttttcggcga gggccttttt gagtatgacc tgctgccctt cctgtcgtcc 3181 accatcagcc cctactaccg ccagtccctc ttccgcaccg tgctggactc cggcatctct 3241 gaggttcgat ccgaccggga caagttcgtc atcttcctcg atgtgaagca cttctccccg 3301 gaggacctca ccgtgaaggt gcaggacgac tttgtggaga tccacggaaa gcacaacgag 3361 cgccaggacg accacggcta catttcccgt gagttccacc gccgctaccg cctgccgtcc 3421 aacgtggacc agtcggccct ctcttgctcc ctgtctgccg atggcatgct gaccttctgt 3481 ggccccaaga tccagactgg cctggatgcc acccacgccg agcgagccat ccccgtgtcg 3541 cgggaggaga agcccacctc ggctccctcg tcctaagcgg ccgcgaagga tctgcgatcg 3601 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 3661 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 3721 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 3781 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg 3841 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg 3901 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta 3961 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc 4021 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt 4081 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga 4141 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 4201 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 4261 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 4321 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 4381 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 4441 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 4501 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 4561 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 4621 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 4681 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg 4741 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 4801 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 4861 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 4921 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 4981 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5041 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5101 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 5161 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5221 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5281 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct 5341 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 5401 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 5461 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 5521 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 5581 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 5641 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 5701 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 5761 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 5821 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 5881 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 5941 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 6001 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6061 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 6121 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 6181 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 6361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 6421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 6481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 6541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 6601 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 6661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 6721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 6781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 6841 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 6901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7021 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7081 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7141 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7201 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7261 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7321 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7381 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7441 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 7501 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 7561 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 7621 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 7681 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 7741 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 7801 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 7861 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 7921 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 7981 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8041 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8101 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8161 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 8221 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 8281 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 8341 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 8401 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 8461 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 8521 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 8581 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 8641 ccaagctg //