LOCUS Exported 7302 bp ds-DNA circular SYN 03-JAN-2023 DEFINITION Expresses FKBP12 protein in mammalian cells. ACCESSION . VERSION . KEYWORDS pLenti6M FKBP12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7302) TITLE Jun Liu Lab Plasmids 2022 REFERENCE 2 (bases 1 to 7302) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 3, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7302 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 42..345 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 346..549 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 500..520 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 546..570 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 632..641 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 638..961 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL LKLE" CDS 1038..1079 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(1116..1136) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1121..1450 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1301..1436 /label=SV40 ori /note="SV40 origin of replication" promoter 1498..1545 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 1564..1962 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" LTR 2049..2282 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2354..2475 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2391..2410) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2445..2464 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2515..2650 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(2670..2689) /label=T7 /note="T7 promoter, forward primer" promoter complement(2671..2689) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2699..2716) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2699..2715) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2708..2730) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2857..3312 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2944..2963) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3154..3175 /label=F1ori-F /note="F1 origin, forward primer" promoter 3338..3442 /gene="bla" /label=AmpR promoter CDS 3443..4303 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3661..3680) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4474..5062 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4963..4982 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5216..5233 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5350..5371 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5386..5416 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5424..5440 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." primer_bind 5429..5451 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5448..5464 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5448..5464 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5483..5503 /label=T3 /note="T3 promoter, forward primer" promoter 5485..5503 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5531..5757 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5758..5938 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5985..6110 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6603..6836 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7021..7065 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" ORIGIN 1 gtgaacggat ctcgacggta tcgataagct tgggagttcc gcgttacata acttacggta 61 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat 121 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg 181 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac 241 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt 301 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg 361 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc 421 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt 481 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata 541 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac 601 ctccatagaa gacaccgact ctagaggatc cgccaccatg ggagtgcagg tggaaaccat 661 ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac 721 cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa 781 gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag 841 tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca 901 cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga 961 aggtaaaaag aagaaaaaga agtcaaagac aaagtgtgta attatgtaac tcgagtctag 1021 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc 1081 gtaccggtta gtaatgagtt tggaattaat tctgtggaat gtgtgtcagt tagggtgtgg 1141 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 1201 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 1261 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 1321 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 1381 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 1441 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt 1501 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa 1561 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca 1621 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac 1681 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc 1741 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc 1801 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat 1861 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg 1921 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacaatt cgagctcggt 1981 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 2041 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac 2101 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc 2161 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt 2221 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag 2281 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc 2341 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 2401 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 2461 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg 2521 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 2581 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 2641 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac 2701 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc 2761 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc 2821 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa 2881 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 2941 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 3001 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 3061 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 3121 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 3181 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 3241 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 3301 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 3361 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 3421 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 3481 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 3541 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 3601 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 3661 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 3721 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 3781 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 3841 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 3901 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 3961 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 4021 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 4081 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 4141 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 4201 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 4261 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 4321 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 4381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 4441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 4501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 4561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 4621 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 4681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 4741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 4801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 4861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 4921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 4981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 5041 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 5101 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 5161 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 5221 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 5281 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 5341 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct 5401 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta 5461 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct 5521 gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt 5581 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg 5641 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc 5701 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg 5761 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 5821 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 5881 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 5941 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga 6001 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa 6061 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc 6121 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat 6181 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg 6241 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc 6301 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc 6361 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa 6421 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat 6481 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta 6541 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga 6601 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca 6661 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat 6721 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag 6781 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg 6841 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg 6901 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa 6961 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa 7021 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac 7081 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt 7141 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca 7201 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa 7261 gaagaaggtg gagagagaga cagagacaga tccattcgat ta //