LOCUS V019948 10596 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019948 VERSION V019948 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..3523 /codon_start=1 /note="mammalian codon-optimized" /product="Aequorea coerulescens GFP" /transl_table=1 /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" /label="AcGFP1" CDS 3557..5389 /label="Men1(NM_001168489)" /note="Men1(NM_001168489)" /gene="Men1" promoter 5431..5930 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5951..6550 /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 6564..7152 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7359..7992 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8121..8137) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8145..8161 /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(8169..8199) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8214..8235 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8523..9111) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9282..10142) /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(10143..10247) /gene="bla" /label="AmpR promoter" polyA_signal 10295..10429 /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 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg 2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct 2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct 3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt 3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg 3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga 3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa 3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa 3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca 3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca 3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt 3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagatctcg 3541 aggaggaggg ggaagcatgg ggctgaaggc cgcccagaag acgctgttcc ctctgcgctc 3601 tatcgacgac gtggtgcgcc tgtttgctgc agagctgggc cgagaggagc ctgacctggt 3661 gctcctgtcc ttggtcctgg gcttcgtgga gcatttcctg gctgtcaacc gtgtcatccc 3721 caccaacgtg ccggagctca ccttccagcc cagccccgca cccgaccctc ctggtggcct 3781 cacctacttc ccggtggccg acctatccat cattgctgcc ctctatgccc gattcaccgc 3841 tcagatccgc ggcgctgtgg acctctccct ctatcctcga gagggaggtg tttctagtcg 3901 cgaactggta aaaaaggtct cggatgtcat atggaacagc ctcagccgct cctacttcaa 3961 ggaccgggcc cacatccagt ccctcttcag cttcatcaca ggcaccaaac tggacagctc 4021 gggcgtggcc tttgcagtgg taggggcctg ccaggctctg ggtctcagag atgtccatct 4081 ggccctgtct gaagatcatg cttgggtggt gtttgggccc aacggggagc agacagctga 4141 ggtgacgtgg cacggcaaag gcaacgagga ccgcagaggc cagacagtca atgccggtgt 4201 ggctgagcgg agctggctgt acctgaaggg gtcgtacatg cgctgcgacc gtaagatgga 4261 ggtggcgttc atggtgtgtg ccatcaaccc ttccatcgat cttcacactg actctttgga 4321 actgttgcaa ctacagcaga agctgctctg gctgctgtat gacctcggac atctggaaag 4381 ataccccatg gcgctaggga acttggcaga cctggaggag ctggagccta ccccgggccg 4441 gccagaccca ctcacccttt atcacaaggg aattgcctca gctaagacct actaccagga 4501 tgaacacatc tacccctaca tgtacctggc tggctaccac tgtcgcaacc gaaatgtgcg 4561 cgaagccctg caggcctggg ccgacactgc cactgttatc caagactaca actactgccg 4621 ggaggatgag gagatctaca aggaattctt tgaagtggcc aatgacgtca tccccaacct 4681 gctgaaggag gcggccagcc tgctagaaac aggcgaagag cggactgggg agcaagccca 4741 gggcacccag ggccaaggtt ctgctctcca ggacccagaa tgctttgccc acctgctgcg 4801 attttatgat ggcatctgca aatgggagga gggtagcccc acaccagtac tgcacgtagg 4861 ctgggcaact ttccttgtgc agtccctagg ccgcttcgag ggacaggtgc ggcagaaggt 4921 gcacatagtt agccgggaag cagaggcggc cgaggctgaa gaaccatggg gggatgaagc 4981 ccgagaaggc cgtcggcgtg gtccccgaag agagtccaag cctgaggagc caccaccacc 5041 caagaagcct gcattggaca agggcccggg ctcaggacaa agtgcagggt cgggaccacc 5101 taggaaaacg tcagggactg tcccaggtac tacccgcggt ggccaggaag tcggcaatgc 5161 tgctcaggct ccagcacctg cagcatcgcc acccccagag ggcccagtgc tcactttcca 5221 gagtgagaag atgaaaggca tgaaggagct actggtggcc accaagatca actcgagcgc 5281 catcaagctg caactcacgg cacagtcgca agtgcagatg aagaaacaga aagtgtccac 5341 acccagcgac tacacactct ctttcctaaa gcggcagcgc aagggcctct gaggatccac 5401 cggatctaga taactgatca taattctacc gggtagggga ggcgcttttc ccaaggcagt 5461 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc 5521 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct 5581 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt 5641 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca 5701 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct 5761 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc 5821 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa 5881 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc 5941 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc 6001 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt 6061 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt 6121 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac 6181 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga 6241 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg 6301 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa 6361 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc 6421 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac 6481 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc 6541 cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga 6601 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 6661 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 6721 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 6781 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 6841 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 6901 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc 6961 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct 7021 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 7081 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 7141 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac 7201 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga 7261 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc 7321 tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca 7381 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta 7441 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa 7501 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt 7561 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag 7621 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa 7681 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg 7741 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt 7801 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac 7861 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg 7921 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga 7981 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga 8041 aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc gacctctagc 8101 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 8161 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 8221 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 8281 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 8341 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 8401 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 8461 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 8521 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 8581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 8641 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 8701 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 8761 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 8821 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 8881 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 8941 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 9001 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 9061 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 9121 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 9181 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 9241 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 9301 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 9361 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 9421 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 9481 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 9541 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 9601 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 9661 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 9721 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 9781 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 9841 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 9901 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 9961 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 10021 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 10081 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 10141 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 10201 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 10261 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg 10321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 10381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac 10441 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta 10501 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa 10561 ttgtatttgt taaatatgta ctacaaactt agtagt //