LOCUS V025935 10688 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025935 VERSION V025935 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" promoter 2337..3519 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" CDS 3551..3568 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 3569..5347 /label="PTH1R(NM_000316)" /note="PTH1R(NM_000316)" /gene="PTH1R" CDS 5354..5419 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 5438..6024 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 6044..6643 /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 6657..7245 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7452..8085 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8213..8229) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8237..8253 /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(8261..8291) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8306..8327 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8615..9203) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9374..10234) /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(10235..10339) /gene="bla" /label="AmpR promoter" polyA_signal 10387..10521 /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt 3541 cgccaccatg caccatcacc atcaccatat ggggaccgcc cggatcgcac ccggcctggc 3601 gctcctgctc tgctgccccg tgctcagctc cgcgtacgcg ctggtggatg cagatgacgt 3661 catgactaaa gaggaacaga tcttcctgct gcaccgtgct caggcccagt gcgaaaaacg 3721 gctcaaggag gtcctgcaga ggccagccag cataatggaa tcagacaagg gatggacatc 3781 tgcgtccaca tcagggaagc ccaggaaaga taaggcatct gggaagctct accctgagtc 3841 tgaggaggac aaggaggcac ccactggcag caggtaccga gggcgcccct gtctgccgga 3901 atgggaccac atcctgtgct ggccgctggg ggcaccaggt gaggtggtgg ctgtgccctg 3961 tccggactac atttatgact tcaatcacaa aggccatgcc taccgacgct gtgaccgcaa 4021 tggcagctgg gagctggtgc ctgggcacaa caggacgtgg gccaactaca gcgagtgtgt 4081 caaatttctc accaatgaga ctcgtgaacg ggaggtgttt gaccgcctgg gcatgattta 4141 caccgtgggc tactccgtgt ccctggcgtc cctcaccgta gctgtgctca tcctggccta 4201 ctttaggcgg ctgcactgca cgcgcaacta catccacatg cacctgttcc tgtccttcat 4261 gctgcgcgcc gtgagcatct tcgtcaagga cgctgtgctc tactctggcg ccacgcttga 4321 tgaggctgag cgcctcaccg aggaggagct gcgcgccatc gcccaggcgc ccccgccgcc 4381 tgccaccgcc gctgccggct acgcgggctg cagggtggct gtgaccttct tcctttactt 4441 cctggccacc aactactact ggattctggt ggaggggctg tacctgcaca gcctcatctt 4501 catggccttc ttctcagaga agaagtacct gtggggcttc acagtcttcg gctggggtct 4561 gcccgctgtc ttcgtggctg tgtgggtcag tgtcagagct accctggcca acaccgggtg 4621 ctgggacttg agctccggga acaaaaagtg gatcatccag gtgcccatcc tggcctccat 4681 tgtgctcaac ttcatcctct tcatcaatat cgtccgggtg ctcgccacca agctgcggga 4741 gaccaacgcc ggccggtgtg acacacggca gcagtaccgg aagctgctca aatccacgct 4801 ggtgctcatg cccctctttg gcgtccacta cattgtcttc atggccacac catacaccga 4861 ggtctcaggg acgctctggc aagtccagat gcactatgag atgctcttca actccttcca 4921 gggatttttt gtcgcaatca tatactgttt ctgcaatggc gaggtacaag ctgagatcaa 4981 gaaatcttgg agccgctgga cactggcact ggacttcaag cgaaaggcac gcagcgggag 5041 cagcagctat agctacggcc ccatggtgtc ccacacaagt gtgaccaatg tcggcccccg 5101 tgtgggactc ggcctgcccc tcagcccccg cctactgccc actgccacca ccaacggcca 5161 ccctcagctg cctggccatg ccaagccagg gaccccagcc ctggagaccc tcgagaccac 5221 accacctgcc atggctgctc ccaaggacga tgggttcctc aacggctcct gctcaggcct 5281 ggacgaggag gcctctgggc ctgagcggcc acctgccctg ctacaggaag agtgggagac 5341 agtcatgtct agagactaca aagaccatga cggtgattat aaagatcatg acatcgacta 5401 caaggatgac gatgacaagt agcggccgcg gatcccgccc ctctccctcc ccccccccta 5461 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt 5521 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga 5581 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg 5641 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt 5701 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat 5761 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg 5821 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg 5881 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt 5941 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa 6001 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc 6061 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc 6121 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt 6181 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 6241 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 6301 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 6361 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 6421 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 6481 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 6541 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 6601 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc 6661 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 6721 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 6781 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 6841 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 6901 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 6961 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 7021 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 7081 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 7141 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 7201 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg 7261 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 7321 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 7381 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 7441 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 7501 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 7561 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 7621 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 7681 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 7741 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 7801 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 7861 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 7921 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 7981 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 8041 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 8101 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt 8161 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 8221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 8281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 8341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 8401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 8461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 8521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 8581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 8641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 8701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 8761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 8821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 8881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 8941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 9001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 9061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 9121 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 9181 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 9241 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 9301 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 9361 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 9421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 9481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 9541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 9601 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 9661 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 9721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 9781 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 9841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 9901 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 9961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 10021 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 10081 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 10141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 10201 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 10261 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 10321 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 10381 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 10441 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 10501 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 10561 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 10621 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 10681 ttagtagt //