LOCUS V031833 10378 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031833 VERSION V031833 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 3572..5140 /label="Rcor2(NM_001320554)" /note="Rcor2(NM_001320554)" /gene="Rcor2" promoter 5213..5712 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5733..6332 /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 6346..6934 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7141..7774 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7903..7919) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7927..7943 /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(7951..7981) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7996..8017 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8305..8893) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9064..9924) /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(9925..10029) /gene="bla" /label="AmpR promoter" polyA_signal 10077..10211 /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 tcagaggagg 3541 tggaggtagt ggaggaggcg gaagcctcga gatgccctca gtaatggaga agcccagtgc 3601 gggctccggg atcttgtccc gaagtcgggc caagacggca cccaacggcg gacagccaca 3661 ctcggaggat gacagcagcg aggaggagca ctctcatgac agcatgatcc gagttggaac 3721 caattaccaa gcagtaattc cggagtgcaa gcctgagagc cctgcacgct atagcaacaa 3781 ggagctgaag ggaatgctgg tgtggtcccc caaccactgt gtatcagatg ccaaacttga 3841 caagtacatt gcaatggcca aggagaagca tggctataac attgagcagg cactcggcat 3901 gcttctgtgg cataaacacg atgtggagaa atcactggct gacctagcca acttcactcc 3961 cttccctgat gagtggacgg tggaggacaa ggtgctgttt gaacaggcct ttgggttcca 4021 tggcaagtgc ttccagcgga tccagcagat gctccctgac aaggtgattc ccagcctggt 4081 gaagtattac tactcttgga agaagacccg aagtcgaact agtgtgatgg acaggcaggc 4141 caggcggctg ggtggccgca aggacaaaga agacagtgat gaactagaag agggacgagg 4201 agctgtgagt gagggggaac cggatactgg agaccccaag cgagagcctc tgccatccag 4261 acctctgaat gcacggcctg gtcctgggaa gaaggaggtt cagatatctc agtatcgcca 4321 ccacccactt cgaactcgga gacgtccgcc caagggcatg tacctgagtc ctgaaggcct 4381 cactgcagta tcgggaagcc cagacctcgc taacctcaca ctccgaggtc ttgactcaca 4441 gctcatctct cttaagcgcc aggtccagag catgaagcag accaatagca gcctccgcca 4501 agcactggag ggcggcattg accctctccg cccccctgag gctaatacca agttcaactc 4561 ccgctggacc actgatgagc aacttctagc tgttcaagct attcgtagat acggcaaaga 4621 ctttggggct attgcagagg tgattgggaa caagactctg acccaagtga agaccttctt 4681 tgtgagctat cgtcgccgct tcaacctgga agaggtgctg caggaatggg aggctgagca 4741 ggatggggcc cccgcagctc cggtccccac ggaagaggcc aggcgagggg ctcctgtacc 4801 agccacagcc ctagaagaag atgatgaggt ccagattaca tctgtctcaa catcagtacc 4861 ccgatcagta ccccctgcac cgcccccacc tccaccgccc acctcgcttt cccagccacc 4921 cccgctgctg aggccacctc tgcccacagc tcccacattg ctccgccagc cgcctcccct 4981 gcagcagggc cgcttcctgc agcctcggct ggcccccaac cagcctccac caccgctcat 5041 ccgccctgcc ctggctgcct ctcgccacag tgcccgccct ggtcctcagc ctccacctac 5101 cctggttgga gctcctttgg agccccctgc accctcactt tgagaattct gcagtcgacg 5161 gtaccgcggg cccgggatcc accggatcta gataactgat cataattcta ccgggtaggg 5221 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg 5281 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg 5341 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc 5401 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact 5461 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc 5521 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc 5581 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc 5641 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct 5701 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc 5761 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact 5821 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc 5881 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 5941 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 6001 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 6061 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 6121 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 6181 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 6241 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 6301 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat 6361 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 6421 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 6481 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6541 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6601 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6661 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6721 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 6781 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6841 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6901 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 6961 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 7021 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 7081 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 7141 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 7201 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 7261 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 7321 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 7381 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 7441 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 7501 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 7561 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 7621 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 7681 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 7741 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 7801 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 7861 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 7921 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 7981 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 8041 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 8101 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 8161 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 8221 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 8281 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 8341 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 8401 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 8461 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 8521 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 8581 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 8641 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 8701 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 8761 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 8821 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 8881 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 8941 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 9001 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 9061 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 9121 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 9181 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 9241 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 9301 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 9361 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 9421 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 9481 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 9541 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 9601 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 9661 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 9721 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 9781 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 9841 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 9901 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 9961 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 10021 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact 10081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 10141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 10201 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat 10261 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg 10321 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt //