LOCUS V024547 9937 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024547 VERSION V024547 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..4705 /label="Cxcr2(NM_009909)" /note="Cxcr2(NM_009909)" /gene="Cxcr2" promoter 4725..5224 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5245..5844 /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 5858..6446 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6653..7286 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7415..7431) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7439..7455 /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(7463..7493) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7508..7529 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7817..8405) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8576..9436) /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(9437..9541) /gene="bla" /label="AmpR promoter" polyA_signal 9589..9723 /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 gggagaattc aaggtggata agttcaacat 3661 tgaagatttc ttcagtggag atcttgatat tttcaattat agctctggca tgccctctat 3721 tctgccagat gctgtcccat gccactcaga gaacctggaa atcaacagtt atgctgtggt 3781 tgtaatatac gtcctggtga ctctgctgag ccttgtgggg aactccttgg tgatgctggt 3841 catcttatac aaccggagca cctgctctgt caccgatgtc tacctgctga acctggccat 3901 tgctgacctg ttctttgccc tgaccttgcc tgtctgggct gcatctaaag taaatggatg 3961 gacttttggc tcaaccctgt gcaagatatt ctcatacgtg aaggaggtta ccttctacag 4021 cagtgttctg ctactagcct gcatcagcat ggaccgctac ctggccattg tacatgccac 4081 aagtacactg atccagaaga gacacttggt caagtttgtg tgcatagcca tgtggttact 4141 atcagtaatt ctggccctgc ccatcttaat tctacgaaat cctgttaagg taaacctttc 4201 taccttagtc tgctatgagg atgtaggtaa caatacatcc cgtttgaggg tcgtactgcg 4261 tatcctgcct cagacttttg gcttcctcgt gccgctgctc atcatgctgt tctgctacgg 4321 gttcacactg cgcaccctct ttaaggccca catggggcag aagcaccggg ccatgcgggt 4381 catcttcgct gtcgtccttg tcttcctgct ctgctggctg ccctacaacc tggttctgtt 4441 cacagacacc ctcatgagaa ccaagctgat caaggagacc tgtgagcgcc gcgatgacat 4501 tgacaaggcc ttgaatgcta cggagattct tggcttcctc cacagctgcc ttaaccccat 4561 catctatgcc tttattggcc agaaatttcg ccatggactt ctcaagatca tggctactta 4621 tggccttgtc agcaaggagt tcttagccaa ggagggaagg ccttcttttg ttagctcgtc 4681 ttcagcaaac acctctacta ccctctaatc tagataattc taccgggtag gggaggcgct 4741 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac 4801 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt 4861 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc 4921 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt 4981 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata 5041 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg 5101 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt 5161 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct 5221 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc 5281 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5341 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5401 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5461 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5521 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5581 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5641 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5701 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5761 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5821 atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg attacaaaat 5881 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 5941 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 6001 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 6061 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 6121 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 6181 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 6241 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct 6301 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 6361 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 6421 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa 6481 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa 6541 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg 6601 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg 6661 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc 6721 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt 6781 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga 6841 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa 6901 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat 6961 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac 7021 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata 7081 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga 7141 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 7201 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 7261 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt 7321 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac 7381 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 7441 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 7501 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 7561 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 7621 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 7681 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 7741 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 7801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 7861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 7921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 7981 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 8041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 8101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 8161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 8221 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 8281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 8341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 8401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 8461 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 8521 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 8581 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 8641 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 8701 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 8761 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 8821 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 8881 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 8941 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 9001 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 9061 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 9121 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 9181 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 9241 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 9301 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 9361 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 9421 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 9481 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 9541 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt 9601 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 9661 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 9721 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt 9781 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt 9841 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa 9901 attgtatttg ttaaatatgt actacaaact tagtagt //