LOCUS V037500 9961 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037500 VERSION V037500 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 2843..3559 /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 3629..4729 /label="Ccr6(NM_009835)" /note="Ccr6(NM_009835)" /gene="Ccr6" promoter 4749..5248 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5269..5868 /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 5882..6470 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6677..7310 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7439..7455) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7463..7479 /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(7487..7517) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7532..7553 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7841..8429) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8600..9460) /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(9461..9565) /gene="bla" /label="AmpR promoter" polyA_signal 9613..9747 /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 tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 3541 catggacgag ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg 3601 gggaagcgga ggcggaggga gtggatccat gaattccaca gagtcctact ttggaacgga 3661 tgattatgac aacacagagt attattctat tcctccagac catgggccat gctccctaga 3721 agaggtcaga aacttcacca aggtatttgt gccaattgcc tactccttaa tatgtgtctt 3781 tggcctcctg ggcaacatta tggtggtgat gacctttgcc ttctacaaga aagccagatc 3841 catgactgac gtctacctgt tgaacatggc catcacagac atactctttg tcctcaccct 3901 accgttctgg gcagttactc atgccaccaa cacttgggtt ttcagcgatg cactgtgtaa 3961 actgatgaaa ggcacatatg cggtcaactt taactgtggg atgctgctcc tggcctgtat 4021 cagcatggac cggtacattg ccatcgtcca ggcaaccaaa tctttccggg tacgctccag 4081 aacactgacg cacagtaagg tcatctgtgt ggcagtgtgg ttcatctcca tcatcatctc 4141 aagccctaca tttatcttca acaagaaata cgagctgcag gatcgtgatg tctgtgagcc 4201 acggtacagg tctgtctcag agcccatcac gtggaagctg ctgggtatgg gactggagct 4261 gttctttggg ttcttcaccc ctttgctgtt tatggtgttc tgctatctgt tcattatcaa 4321 gaccttggtg caggcccaga actccaagag gcacagagca atccgagtcg tgatcgctgt 4381 ggttctcgtg ttcctggctt gtcagatccc tcacaacatg gtcctcctcg tgactgcggt 4441 caacacgggc aaagtgggcc ggagctgcag caccgagaaa gtcctcgcct acaccaggaa 4501 cgtggccgag gtcctggctt tcctgcattg ctgcctcaac cccgtgttgt atgcgtttat 4561 tggacagaaa ttcagaaact acttcatgaa gatcatgaag gatgtgtggt gtatgagaag 4621 gaagaataag atgcctggct tcctctgtgc ccgggtttac tcggaaagct acatctccag 4681 gcagaccagt gagaccgtcg aaaatgataa tgcatcgtcc tttaccatgt aatctagata 4741 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc 4801 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac 4861 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc 4921 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga 4981 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt 5041 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct 5101 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg 5161 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt 5221 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac 5281 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5341 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5401 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5461 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5521 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5581 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5641 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5701 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5761 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5821 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa 5881 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5941 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 6001 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 6061 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 6121 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 6181 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 6241 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 6301 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 6361 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6421 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa 6481 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc 6541 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca 6601 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 6661 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat 6721 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca 6781 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat 6841 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat 6901 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt 6961 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc 7021 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag 7081 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg 7141 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 7201 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 7261 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 7321 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 7381 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt 7441 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7501 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7561 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7621 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 8041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 8101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 8161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 8221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 8281 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 8341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 8401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 9001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 9061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 9121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 9181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 9241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 9301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 9361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 9601 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 9661 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 9721 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc 9781 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa 9841 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact 9901 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag 9961 t //