LOCUS V043887 10255 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043887 VERSION V043887 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2815..3531 /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 3553..4947 /label="PRKN(NM_004562)" /note="PRKN(NM_004562)" /gene="PRKN" misc_feature 4958..5544 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5564..6163 /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 6177..6765 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6972..7605 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7733..7749) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7757..7773 /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(7781..7811) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7826..7847 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8135..8723) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8894..9754) /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(9755..9859) /gene="bla" /label="AmpR promoter" polyA_signal 9907..10041 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggtg 2821 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 2881 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 2941 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 3001 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 3061 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 3121 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 3181 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 3241 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 3301 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 3361 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 3421 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 3481 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggcgga 3541 gggagtctcg agatgatagt gtttgtcagg ttcaactcca gccatggttt cccagtggag 3601 gtcgattctg acaccagcat cttccagctc aaggaggtgg ttgctaagcg acagggggtt 3661 ccggctgacc agttgcgtgt gattttcgca gggaaggagc tgaggaatga ctggactgtg 3721 cagaattgtg acctggatca gcagagcatt gttcacattg tgcagagacc gtggagaaaa 3781 ggtcaagaaa tgaatgcaac tggaggcgac gaccccagaa acgcggcggg aggctgtgag 3841 cgggagcccc agagcttgac tcgggtggac ctcagcagct cagtcctccc aggagactct 3901 gtggggctgg ctgtcattct gcacactgac agcaggaagg actcaccacc agctggaagt 3961 ccagcaggta gatcaatcta caacagcttt tatgtgtatt gcaaaggccc ctgtcaaaga 4021 gtgcagccgg gaaaactcag ggtacagtgc agcacctgca ggcaggcaac gctcaccttg 4081 acccagggtc catcttgctg ggatgatgtt ttaattccaa accggatgag tggtgaatgc 4141 caatccccac actgccctgg gactagtgca gaatttttct ttaaatgtgg agcacacccc 4201 acctctgaca aggaaacatc agtagctttg cacctgatcg caacaaatag tcggaacatc 4261 acttgcatta cgtgcacaga cgtcaggagc cccgtcctgg ttttccagtg caactcccgc 4321 cacgtgattt gcttagactg tttccactta tactgtgtga caagactcaa tgatcggcag 4381 tttgttcacg accctcaact tggctactcc ctgccttgtg tggctggctg tcccaactcc 4441 ttgattaaag agctccatca cttcaggatt ctgggagaag agcagtacaa ccggtaccag 4501 cagtatggtg cagaggagtg tgtcctgcag atggggggcg tgttatgccc ccgccctggc 4561 tgtggagcgg ggctgctgcc ggagcctgac cagaggaaag tcacctgcga agggggcaat 4621 ggcctgggct gtgggtttgc cttctgccgg gaatgtaaag aagcgtacca tgaaggggag 4681 tgcagtgccg tatttgaagc ctcaggaaca actactcagg cctacagagt cgatgaaaga 4741 gccgccgagc aggctcgttg ggaagcagcc tccaaagaaa ccatcaagaa aaccaccaag 4801 ccctgtcccc gctgccatgt accagtggaa aaaaatggag gctgcatgca catgaagtgt 4861 ccgcagcccc agtgcaggct cgagtggtgc tggaactgtg gctgcgagtg gaaccgcgtc 4921 tgcatggggg accactggtt cgacgtgtag gatcccgccc ctctccctcc ccccccccta 4981 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt 5041 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga 5101 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg 5161 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt 5221 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat 5281 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg 5341 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg 5401 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt 5461 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa 5521 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc 5581 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc 5641 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt 5701 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 5761 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 5821 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 5881 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 5941 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 6001 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 6061 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 6121 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc 6181 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 6241 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 6301 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 6361 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 6421 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 6481 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 6541 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 6601 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 6661 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 6721 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg 6781 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 6841 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 6901 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6961 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 7021 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 7081 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 7141 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 7201 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 7261 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 7321 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 7381 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 7441 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 7501 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 7561 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 7621 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt 7681 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 7741 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7801 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7921 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7981 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 8041 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 8101 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 8161 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 8221 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 8281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 8341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 8401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 8461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 8521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 8581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8701 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8821 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8881 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8941 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 9001 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 9061 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 9121 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 9181 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 9241 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 9301 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 9361 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 9421 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 9481 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 9541 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 9601 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 9661 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9721 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9781 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9841 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 9901 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 10021 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 10081 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 10141 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 10201 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //