LOCUS V022173 9468 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022173 VERSION V022173 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 2807..3514 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 3548..4261 /label="Cd63(NM_007653)" /note="Cd63(NM_007653)" /gene="Cd63" promoter 4303..4802 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4823..5422 /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 5436..6024 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6231..6864 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6993..7009) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7017..7033 /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(7041..7071) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7086..7107 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7395..7983) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8154..9014) /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(9015..9119) /gene="bla" /label="AmpR promoter" polyA_signal 9167..9301 /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 tctaccggtc gccaccatgg tgagcaaggg 2821 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg 2881 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg 2941 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat 3001 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc 3061 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga 3121 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta 3181 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac 3241 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga 3301 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac 3361 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga 3421 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg 3481 ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc gaggaggagg 3541 tggaagtatg gcggtggaag gaggaatgaa gtgtgtcaag tttttgctct acgttctcct 3601 gctggccttc tgcgcctgtg cagtgggatt gatcgccatt ggtgtagcgg ttcaggttgt 3661 cttgaagcag gccattaccc atgagactac tgctggctcg ctgttgcctg tggtcatcat 3721 tgcagtgggt gccttcctct tcctggtggc ctttgtgggc tgctgtgggg cctgcaagga 3781 gaactactgt ctcatgatta catttgccat cttcctgtct cttatcatgc ttgtggaggt 3841 ggctgtggcc attgctggct atgtgtttag agaccaggtg aagtcagagt ttaataaaag 3901 cttccagcag cagatgcaga attaccttaa agacaacaaa acagccacta ttttggacaa 3961 attgcagaaa gaaaataact gctgtggagc ttctaactac acagactggg aaaacatccc 4021 cggcatggcc aaggacagag tccccgattc ttgctgcatc aacataactg tgggctgtgg 4081 gaatgatttc aaggaatcca ctatccatac ccagggctgc gtggagacta tagcaatatg 4141 gctaaggaag aacatactgc tggtggctgc agcggccctg ggcattgctt ttgtggaggt 4201 cttgggaatt atcttctcct gctgtctggt gaagagtatt cgaagtggct atgaagtaat 4261 gtagggatcc accggatcta gataactgat cataattcta ccgggtaggg gaggcgcttt 4321 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa 4381 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct 4441 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc 4501 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc 4561 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc 4621 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc 4681 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct 4741 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt 4801 cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg 4861 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 4921 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt 4981 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 5041 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 5101 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 5161 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga 5221 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 5281 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 5341 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 5401 gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat tacaaaattt 5461 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 5521 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 5581 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 5641 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 5701 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 5761 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 5821 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc 5881 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 5941 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 6001 tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca 6061 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 6121 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 6181 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct 6241 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 6301 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg 6361 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga 6421 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 6481 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc 6541 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt 6601 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata 6661 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc 6721 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 6781 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 6841 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat 6901 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg 6961 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 7021 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 7081 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 7141 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 7201 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 7261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 7321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 7381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 7441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 7501 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 7561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 7621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 7681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 7741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 7801 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 7861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 7921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 7981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 8041 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 8101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 8161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 8221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 8281 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 8341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 8401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 8461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 8521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 8581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 8641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 8701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 8761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 8821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 8881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 8941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 9001 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 9061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 9121 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc 9181 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 9241 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat 9301 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac 9361 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc 9421 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt //