LOCUS Exported 6508 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION MESA split-TEV protease chain with Rapamycin FKBP ectodomain, 2 aa scaffold, 6 aa linker domain, N-terminal TEV protease. ACCESSION . VERSION . KEYWORDS FKBP_SCF2_LD6_NTEV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6508) AUTHORS Daringer NM, Dudek RM, Schwarz KA, Leonard JN TITLE Modular Extracellular Sensor Architecture for Engineering Mammalian Cell-based Devices. JOURNAL ACS Synth Biol. 2014 Mar 11. PUBMED 24611683 REFERENCE 2 (bases 1 to 6508) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6508 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 384..587 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 538..558 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 632..651 /label=T7 /note="T7 promoter, forward primer" promoter 632..650 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 754..1077 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL LKLE" misc_feature 1139..1191 /label=MCS /note="multiple cloning site" polyA_signal 1784..2176 /label=-beta-globin poly(A) /note="human beta-globin polyadenylation signal" rep_origin 3084..3597 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3171..3190) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3381..3402 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4078..4097) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4197..4219 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4257..4275) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4343..4447 /gene="bla" /label=AmpR promoter CDS 4448..5308 /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(4666..4685) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5479..6067 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5968..5987 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6221..6238 /label=L4440 /note="L4440 vector, forward primer" repeat_region 6307..6436 /label=AAV2 ITR ORIGIN 1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 661 ctggctagcg ccatcatgtg ccgagccatc tctcttaggc gcttgctgct gctgctgctg 721 cagctgtcac aactcctagc tgtcactcaa gggatgggag tgcaggtgga aaccatctcc 781 ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg 841 atgcttgaag atggaaagaa atttgattcc tcccgggaca gaaacaagcc ctttaagttt 901 atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg 961 ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca 1021 ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaagt 1081 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtcct 1141 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct 1201 agaaccggta agcttttttg ggcactggtc gtggttgctg gagtcctgtt ttgttatggc 1261 ttgctagtga cagtggctct ttgtgttatc tgggtaagat ctggtgagag cttgtttaag 1321 gggccgcgtg attacaaccc gatatcgagc accatttgtc atttgacgaa tgaatctgat 1381 gggcacacaa catcgttgta tggtattgga tttggtccct tcatcattac aaacaagcac 1441 ttgtttagaa gaaataatgg aacactgttg gtccaatcac tacatggtgt attcaaggtc 1501 aagaacacca cgactttgca acaacacctc attgatggga gggacatgat aattattcgc 1561 atgcctaagg atttcccacc atttcctcaa aagctgaaat ttagagagcc acaaagggaa 1621 gagcgcatat gtcttgtgac aaccaacttc caaactaaga gctaggcggc cgccaccgcg 1681 gtggagctgt ttaaacctcg agcaattcac cccaccagtg caggctgcct atcagaaagt 1741 ggtggctggt gtggctaatg ccctggccca caagtatcac taagctcgct ttcttgctgt 1801 ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg gggatattat 1861 gaagggcctt gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat 1921 gtatttaaat tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta 1981 aaacataaag aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc 2041 catgaaagaa ggtgaggctg caaacagtaa tgcacattgg caacagcccc tgatgcctat 2101 gccttattca tccctcagaa aaggattcaa gtagaggctt gatttggagg ttaaagtttt 2161 gctatgctgt attttacatt acttattgtt ttagctgtcc tcatgaatgt cttttcacta 2221 cccatttgct tatcctgcat ctctcagcct tgactccact cagttctctt gcttagagat 2281 accacctttc ccctgaagtg ttccttccat gttttacggc gagatggttt ctcctcgcct 2341 ggccactcag ccttagttgt ctctgttgtc ttatagaggt ctacttgaag aaggaaaaac 2401 agggggcatg gtttgactgt cctgtgagcc cttcttccct gcctccccca ctcacagtga 2461 cccggaatcc ctcgacatgg cagtctagag catggctacg tagataagta gcatggcggg 2521 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct 2581 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg 2641 gcctcagtga gcgagcgagc gcgccagctg gcgtaatagc gaagaggccc gcaccgatcg 2701 cccttcccaa cagttgcgca gcctgaatgg cgaatggaat tccagacgat tgagcgtcaa 2761 aatgtaggta tttccatgag cgtttttcct gttgcaatgg ctggcggtaa tattgttctg 2821 gatattacca gcaaggccga tagtttgagt tcttctactc aggcaagtga tgttattact 2881 aatcaaagaa gtattgcgac aacggttaat ttgcgtgatg gacagactct tttactcggt 2941 ggcctcactg attataaaaa cacttctcag gattctggcg taccgttcct gtctaaaatc 3001 cctttaatcg gcctcctgtt tagctcccgc tctgattcta acgaggaaag cacgttatac 3061 gtgctcgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 3121 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 3181 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 3241 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 3301 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3361 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3421 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3481 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta 3541 aatatttgct tatacaatct tcctgttttt ggggcttttc tgattatcaa ccggggtaca 3601 tatgattgac atgctagttt tacgattacc gttcatcgat tctcttgttt gctccagact 3661 ctcaggcaat gacctgatag cctttgtaga gacctctcaa aaatagctac cctctccggc 3721 atgaatttat cagctagaac ggttgaatat catattgatg gtgatttgac tgtctccggc 3781 ctttctcacc cgtttgaatc tttacctaca cattactcag gcattgcatt taaaatatat 3841 gagggttcta aaaattttta tccttgcgtt gaaataaagg cttctcccgc aaaagtatta 3901 cagggtcata atgtttttgg tacaaccgat ttagctttat gctctgaggc tttattgctt 3961 aattttgcta attctttgcc ttgcctgtat gatttattgg atgttggaat tcctgatgcg 4021 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac 4081 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc 4141 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 4201 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct 4261 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg 4321 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 4381 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 4441 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 4501 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 4561 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 4621 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 4681 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 4741 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 4801 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 4861 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 4921 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 4981 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 5041 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 5101 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 5161 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 5221 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 5281 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 5341 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 5401 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 5461 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 5521 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 5581 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 5641 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 5701 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 5761 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 5821 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 5881 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 5941 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 6001 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 6061 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 6121 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 6181 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 6241 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 6301 cagcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg 6361 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc 6421 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca 6481 tgctctaggg atccggccag atatacgc //