LOCUS V022271 9174 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022271 VERSION V022271 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" misc_feature 2811..2867 /note="multiple cloning site" /label="MCS" misc_feature 2869..3455 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 3456..3944 /codon_start=1 /note="This allele of Thy1 is also known as CD90.1." /product="mouse GPI-anchored cell surface glycoprotein of the immunoglobulin superfamily" /transl_table=1 /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL* " /label="Thy1.1" promoter 3962..4461 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4482..5081 /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 5095..5683 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5890..6523 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6652..6668) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6676..6692 /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(6700..6730) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6745..6766 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7054..7642) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7813..8673) /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(8674..8778) /gene="bla" /label="AmpR promoter" polyA_signal 8826..8960 /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 gaattctgca gtcgacggta ccgcgggccc gggatccgcc cctctccctc 2881 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta 2941 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc 3001 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct 3061 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt 3121 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa 3181 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg 3241 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga 3301 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac 3361 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt 3421 cctttgaaaa acacgatgat aatatggcca caaccatgaa cccagccatc agcgtcgctc 3481 tcctgctctc agtcttgcag gtgtcccgag ggcagaaggt gaccagcctg acagcctgcc 3541 tggtgaacca aaaccttcgc ctggactgcc gccatgagaa taacaccaag gataactcca 3601 tccagcatga gttcagcctg acccgagaga agaggaagca cgtgctctca ggcacccttg 3661 ggatacccga gcacacgtac cgctcccgcg tcaccctctc caaccagccc tatatcaagg 3721 tccttaccct agccaacttc accaccaagg atgagggcga ctacttttgt gagcttcgcg 3781 tctcgggcgc gaatcccatg agctccaata aaagtatcag tgtgtataga gacaagctgg 3841 tcaagtgtgg cggcataagc ctgctggttc agaacacatc ctggatgctg ctgctgctgc 3901 tttccctctc cctcctccaa gccctggact tcatttctct gtgaatctag ataattctac 3961 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg 4021 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg 4081 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc 4141 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca 4201 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt 4261 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg 4321 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc 4381 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct 4441 tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca 4501 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 4561 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 4621 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 4681 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 4741 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 4801 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 4861 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 4921 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 4981 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 5041 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa 5101 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5161 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5221 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5281 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5341 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5401 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5461 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 5521 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 5581 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 5641 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca 5701 gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg 5761 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg 5821 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 5881 agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga 5941 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca 6001 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag 6061 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg 6121 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg 6181 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg 6241 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag 6301 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac 6361 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 6421 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 6481 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat 6541 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga 6601 cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 6661 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 6721 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 6781 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 6841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 6901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 6961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 7021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 7081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 7141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 7201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 7261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 7321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 7381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 7441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 7501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 7561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 7621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 7681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 7801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 7861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 7921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 7981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 8041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 8101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 8161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 8221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 8281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 8341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 8401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 8461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 8521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 8581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 8701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 8761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 8821 agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 8881 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 8941 tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc 9001 ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg 9061 attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact 9121 tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //