LOCUS V045777 10209 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045777 VERSION V045777 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..4881 /label="GRK2(NM_001619)" /note="GRK2(NM_001619)" /gene="GRK2" misc_feature 4912..5498 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5518..6117 /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 6131..6719 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6926..7559 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7687..7703) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7711..7727 /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(7735..7765) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7780..7801 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8089..8677) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8848..9708) /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(9709..9813) /gene="bla" /label="AmpR promoter" polyA_signal 9861..9995 /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 caccatggcg 2821 gacctggagg cggtgctggc cgacgtgagc tacctgatgg ccatggagaa gagcaaggcc 2881 acgccggccg cgcgcgccag caagaagatc ctgctgcccg agcccagcat ccgcagtgtc 2941 atgcagaagt acctggagga ccggggcgag gtgacctttg agaagatctt ttcccagaag 3001 ctggggtacc tgctcttccg agacttctgc ctgaaccacc tggaggaggc caggcccttg 3061 gtggaattct atgaggagat caagaagtac gagaagctgg agacggagga ggagcgtgtg 3121 gcccgcagcc gggagatctt cgactcatac atcatgaagg agctgctggc ctgctcgcat 3181 cccttctcga agagtgccac tgagcatgtc caaggccacc tggggaagaa gcaggtgcct 3241 ccggatctct tccagccata catcgaagag atttgtcaaa acctccgagg ggacgtgttc 3301 cagaaattca ttgagagcga taagttcaca cggttttgcc agtggaagaa tgtggagctc 3361 aacatccacc tgaccatgaa tgacttcagc gtgcatcgca tcattgggcg cgggggcttt 3421 ggcgaggtct atgggtgccg gaaggctgac acaggcaaga tgtacgccat gaagtgcctg 3481 gacaaaaagc gcatcaagat gaagcagggg gagaccctgg ccctgaacga gcgcatcatg 3541 ctctcgctcg tcagcactgg ggactgccca ttcattgtct gcatgtcata cgcgttccac 3601 acgccagaca agctcagctt catcctggac ctcatgaacg gtggggacct gcactaccac 3661 ctctcccagc acggggtctt ctcagaggct gacatgcgct tctatgcggc cgagatcatc 3721 ctgggcctgg agcacatgca caaccgcttc gtggtctacc gggacctgaa gccagccaac 3781 atccttctgg acgagcatgg ccacgtgcgg atctcggacc tgggcctggc ctgtgacttc 3841 tccaagaaga agccccatgc cagcgtgggc acccacgggt acatggctcc ggaggtcctg 3901 cagaagggcg tggcctacga cagcagtgcc gactggttct ctctggggtg catgctcttc 3961 aagttgctgc gggggcacag ccccttccgg cagcacaaga ccaaagacaa gcatgagatc 4021 gaccgcatga cgctgacgat ggccgtggag ctgcccgact ccttctcccc tgaactacgc 4081 tccctgctgg aggggttgct gcagagggat gtcaaccgga gattgggctg cctgggccga 4141 ggggctcagg aggtgaaaga gagccccttt ttccgctccc tggactggca gatggtcttc 4201 ttgcagaagt accctccccc gctgatcccc ccacgagggg aggtgaacgc ggccgacgcc 4261 ttcgacattg gctccttcga tgaggaggac acaaaaggaa tcaagttact ggacagtgat 4321 caggagctct accgcaactt ccccctcacc atctcggagc ggtggcagca ggaggtggca 4381 gagactgtct tcgacaccat caacgctgag acagaccggc tggaggctcg caagaaagcc 4441 aagaacaagc agctgggcca tgaggaagac tacgccctgg gcaaggactg catcatgcat 4501 ggctacatgt ccaagatggg caaccccttc ctgacccagt ggcagcggcg gtacttctac 4561 ctgttcccca accgcctcga gtggcggggc gagggcgagg ccccgcagag cctgctgacc 4621 atggaggaga tccagtcggt ggaggagacg cagatcaagg agcgcaagtg cctgctcctc 4681 aagatccgcg gtgggaaaca gttcattttg cagtgcgata gcgaccctga gctggtgcag 4741 tggaagaagg agctgcgcga cgcctaccgc gaggcccagc agctggtgca gcgggtgccc 4801 aagatgaaga acaagccgcg ctcgcccgtg gtggagctga gcaaggtgcc gctggtccag 4861 cgcggcagtg ccaacggcct ctaactagtt ctagagcggc cgcggatccc gcccctctcc 4921 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 4981 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 5041 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 5101 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc 5161 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 5221 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 5281 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa 5341 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 5401 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt 5461 tttcctttga aaaacacgat gataagcttg ccacaaccca caaggagacg accttccatg 5521 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc 5581 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc 5641 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5701 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5761 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5821 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5881 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5941 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 6001 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 6061 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctagacg 6121 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 6181 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 6241 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 6301 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 6361 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 6421 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 6481 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 6541 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 6601 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 6661 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 6721 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 6781 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 6841 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 6901 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 6961 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 7021 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 7081 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 7141 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 7201 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 7261 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 7321 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 7381 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 7441 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 7501 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 7561 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 7621 gagtgagagg ccttgacatt gctagcgttt accgtcgacc tctagctaga gcttggcgta 7681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 7741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 7801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 7861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 7921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 7981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 8041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 8101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 8161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 8221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 8281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 8341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 8401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 8461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 8521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 8581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 8641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 8701 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 8761 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 8821 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 8881 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 8941 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 9001 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 9061 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 9121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 9181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 9241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 9301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 9361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 9421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 9481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 9541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 9601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 9661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 9721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 9781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 9841 cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca 9901 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 9961 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa 10021 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt 10081 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa 10141 tatgtactac aaacttagta gtttttaaag aaattgtatt tgttaaatat gtactacaaa 10201 cttagtagt //