LOCUS       sequence-138316-        7614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA protease chain with Rapamycin FKBP ectodomain and no linker 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7614)
  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 7614)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2014 Mar 11."
COMMENT     Sequence Label: FKBP_LD 0_TEV
FEATURES             Location/Qualifiers
     source          1..7614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        555..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     709..729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     803..822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        803..821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             925..951
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(958..960,961..1281)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    1343..1395
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1492..2196
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     CDS             2230..2940
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2375..2393)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2643..2662
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3061..3453
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      4361..4874
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4448..4467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4658..4679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5355..5374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5474..5496
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5534..5552)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5620..5724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5725..5793,5794..6585)
                     /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(5943..5962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6756..7344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7245..7264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7584..7614,1..99)
                     /label=AAV2 ITR
ORIGIN
        1 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
       61 gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc
      121 catgctactt atctacgtag ccatgctcta gggatccggc cagatatacg cgttgacatt
      181 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      541 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      721 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      781 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      841 gccatcatgt gccgagccat ctctcttagg cgcttgctgc tgctgctgct gcagctgtca
      901 caactcctag ctgtcactca agggtacccc tatgatgtgc cagactacgc cggtaccatg
      961 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag
     1021 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
     1081 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
     1141 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
     1201 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
     1261 gtggagcttc taaaactgga aagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     1321 gctggtggta gtgctggtgg tcctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcgggcccgg gatccaccgg atctagaacc ggtaagcttt tttgggcact ggtcgtggtt
     1441 gctggagtcc tgttttgtta tggcttgcta gtgacagtgg ctctttgtgt tgagagcttg
     1501 tttaaggggc cgcgtgatta caacccgata tcgagcacca tttgtcattt gacgaatgaa
     1561 tctgatgggc acacaacatc gttgtatggt attggatttg gtcccttcat cattacaaac
     1621 aagcacttgt ttagaagaaa taatggaaca ctgttggtcc aatcactaca tggtgtattc
     1681 aaggtcaaga acaccacgac tttgcaacaa cacctcattg atgggaggga catgataatt
     1741 attcgcatgc ctaaggattt cccaccattt cctcaaaagc tgaaatttag agagccacaa
     1801 agggaagagc gcatatgtct tgtgacaacc aacttccaaa ctaagagcat gtctagcatg
     1861 gtgtcagaca ctagttgcac attcccttca tctgatggca tattctggaa gcattggatt
     1921 caaaccaagg atgggcagtg tggcagtcca ttagtatcaa ctagagatgg gttcattgtt
     1981 ggtatacact cagcatcgaa tttcaccaac acaaacaatt atttcacaag cgtgccgaaa
     2041 aacttcatgg aattgttgac aaatcaggag gcgcagcagt gggttagtgg ttggcgatta
     2101 aatgctgact cagtattgtg ggggggccat aaagttttca tggtgaaacc tgaagagcct
     2161 tttcagccag ttaaggaagc gactcaactc atgaatggag gtaccggcgg aggctccggt
     2221 ggtggctcta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     2281 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     2341 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     2401 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     2461 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     2521 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     2581 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     2641 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     2701 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     2761 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     2821 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     2881 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     2941 tagcggccgc caccgcggtg gagctgttta aacctcgagc aattcacccc accagtgcag
     3001 gctgcctatc agaaagtggt ggctggtgtg gctaatgccc tggcccacaa gtatcactaa
     3061 gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac
     3121 taaactgggg atattatgaa gggccttgag catctggatt ctgcctaata aaaaacattt
     3181 attttcattg caatgatgta tttaaattat ttctgaatat tttactaaaa agggaatgtg
     3241 ggaggtcagt gcatttaaaa cataaagaaa tgaagagcta gttcaaacct tgggaaaata
     3301 cactatatct taaactccat gaaagaaggt gaggctgcaa acagtaatgc acattggcaa
     3361 cagcccctga tgcctatgcc ttattcatcc ctcagaaaag gattcaagta gaggcttgat
     3421 ttggaggtta aagttttgct atgctgtatt ttacattact tattgtttta gctgtcctca
     3481 tgaatgtctt ttcactaccc atttgcttat cctgcatctc tcagccttga ctccactcag
     3541 ttctcttgct tagagatacc acctttcccc tgaagtgttc cttccatgtt ttacggcgag
     3601 atggtttctc ctcgcctggc cactcagcct tagttgtctc tgttgtctta tagaggtcta
     3661 cttgaagaag gaaaaacagg gggcatggtt tgactgtcct gtgagccctt cttccctgcc
     3721 tcccccactc acagtgaccc ggaatccctc gacatggcag tctagagcat ggctacgtag
     3781 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3841 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3901 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg ccagctggcg taatagcgaa
     3961 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaattcc
     4021 agacgattga gcgtcaaaat gtaggtattt ccatgagcgt ttttcctgtt gcaatggctg
     4081 gcggtaatat tgttctggat attaccagca aggccgatag tttgagttct tctactcagg
     4141 caagtgatgt tattactaat caaagaagta ttgcgacaac ggttaatttg cgtgatggac
     4201 agactctttt actcggtggc ctcactgatt ataaaaacac ttctcaggat tctggcgtac
     4261 cgttcctgtc taaaatccct ttaatcggcc tcctgtttag ctcccgctct gattctaacg
     4321 aggaaagcac gttatacgtg ctcgtcaaag caaccatagt acgcgccctg tagcggcgca
     4381 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4441 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4501 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4561 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4621 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4681 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4741 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4801 ttaacgttta caatttaaat atttgcttat acaatcttcc tgtttttggg gcttttctga
     4861 ttatcaaccg gggtacatat gattgacatg ctagttttac gattaccgtt catcgattct
     4921 cttgtttgct ccagactctc aggcaatgac ctgatagcct ttgtagagac ctctcaaaaa
     4981 tagctaccct ctccggcatg aatttatcag ctagaacggt tgaatatcat attgatggtg
     5041 atttgactgt ctccggcctt tctcacccgt ttgaatcttt acctacacat tactcaggca
     5101 ttgcatttaa aatatatgag ggttctaaaa atttttatcc ttgcgttgaa ataaaggctt
     5161 ctcccgcaaa agtattacag ggtcataatg tttttggtac aaccgattta gctttatgct
     5221 ctgaggcttt attgcttaat tttgctaatt ctttgccttg cctgtatgat ttattggatg
     5281 ttggaattcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5341 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5401 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5461 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5521 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5581 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5641 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5701 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5761 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5821 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5881 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5941 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6001 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6061 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6121 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6181 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6241 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6301 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6361 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6421 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6481 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6541 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6661 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6721 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6841 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6901 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6961 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7021 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7081 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7141 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7201 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7261 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7321 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7381 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7441 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7501 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7561 tggccgattc attaatgcag cagctgcgcg ctcgctcgct cactgaggcc gccc
//
