LOCUS V033900 9825 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033900 VERSION V033900 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..4531 /label="ITPRIPL1(NM_178495)" /note="ITPRIPL1(NM_178495)" /gene="ITPRIPL1" CDS 4532..4555 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 4559..4582 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 4613..5112 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5133..5732 /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 5746..6334 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6541..7174 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7303..7319) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7327..7343 /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(7351..7381) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7396..7417 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7705..8293) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8464..9324) /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(9325..9429) /gene="bla" /label="AmpR promoter" polyA_signal 9477..9611 /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 ccatgaccac tgactcctac tctcctactt ctccagatgc 2881 agaggcctcc atggctgtga taagcctgct gttcttggca gtgatgtatg ttgttcacca 2941 ccctctgatg gtcagtgatc ggatggacct ggacacatta gccaggagtc ggcagctgga 3001 gaagcgaatg agtgaggaga tgcgcctgct agagatggag tttgaagaga gaaagcgagc 3061 cgctgagcag aggcagaagg cagagaactt ctggacagga gacacatcca gtgaccagtt 3121 agtgctgggg aagaaagaca tggggtggcc gttccaggcc gatggccagg aagggcctct 3181 gggctggatg ctgggaaacc tgtggaacac tggcctcttt tgcctttttc tcgtctttga 3241 gctcctgcga cagaacatgc agcatgaacc ggcctttgat tccagcagtg aggaggagga 3301 ggaggaagtc cgtgttgtcc ctgtcacctc ttacaactgg cttactgact tcccctccca 3361 ggaggccctg gactcctttt acaaacacta tgtccaaaat gccatccgtg acctgccctg 3421 cacctgtgag tttgtggaga gttttgtgga tgatctcatt gaggcctgtc gggtgctcag 3481 ccgccaagag gctcacccac aattggaaga ctgcctgggc atcggggctg cctttgagaa 3541 atggggaacc ctccatgaga cccagaaatt tgatatcctg gtgcccattg tccccccaca 3601 gggcaccatg tttgtcctgg agatgaggga cccagccctg ggccgccgct gtggctgtgt 3661 gctggtggag tcagaatgtg tgtgcaagcg tgagaaactc ctaggggacg tgctgtgcct 3721 ggtgcaccac cacagggacc cctcggcagt cttggggaag tgtagtagct ccatcaaggc 3781 agctctctgc accggcttcc acctagacgt gtgcaagact gtgcagtggt tccggaacat 3841 gatgggcaat gcctgggccc ttgtggccca caagtatgac tttaaactca gtctcccacc 3901 gtctaccacc tcctgcaagc tccggctgga ctatcgctca ggccgctttc tctcaatcca 3961 cttggtcctg ggggtgcaac gagaagacac cttggtctac ctggtgagtc aggctcctga 4021 ccaggagcag ctcaccagtg tggactggcc tgagtccttt gtggcctgtg agcacctgtt 4081 cctgaagctg gtggggcgct ttgcccccga gaacacctgt cacctcaagt gcctccagat 4141 cattttaagt ctccggcagc atcagagctt accccatgga gcatcccgcc ccatcctcac 4201 ttcttaccat tttaaaacag ctctcatgca cctcttgcta cggctgcccc tcacggactg 4261 ggcccacaac atgctctctc agcggctcca ggacattctc tggttcttgg gccgtggcct 4321 ccagcaaagg tccctccatc atttcctcat tggtaacaat tttctgcccc tgaccatccc 4381 aatccctaag acatttagga atgctgagcc ggtcaatctc ttccaacacc tggtgctcaa 4441 ccccaaggca cattcacagg cagtggaaga gttccaaaac cttctgaccc aggtgaaaac 4501 tctgcctcat gccccactgg ctgcagcacc tgactacaag gacgacgatg acaaggcaga 4561 ctacaaggac gacgatgaca agtaggatcc cgcgactcta gataattcta ccgggtaggg 4621 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg 4681 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg 4741 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc 4801 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact 4861 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc 4921 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc 4981 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc 5041 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct 5101 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc 5161 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact 5221 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc 5281 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 5341 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 5401 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 5461 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 5521 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 5581 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 5641 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 5701 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat 5761 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5821 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5881 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5941 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6001 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6061 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6121 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 6181 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6241 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6301 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 6361 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 6421 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 6481 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 6541 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 6601 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 6661 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 6721 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 6781 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 6841 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 6901 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 6961 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 7021 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 7081 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 7141 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 7201 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 7261 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 7321 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 7381 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 7441 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 7501 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 7561 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 7621 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 7681 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 7741 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 7801 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 7861 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 7921 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 7981 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 8041 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 8101 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 8161 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 8221 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 8281 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 8341 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 8401 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 8461 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 8521 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 8581 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 8641 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 8701 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 8761 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 8821 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 8881 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 8941 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 9001 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 9061 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 9121 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 9181 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 9241 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 9301 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 9361 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 9421 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact 9481 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 9541 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 9601 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat 9661 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg 9721 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt 9781 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //