LOCUS       sequence-94240-e        6328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA split-TEV protease chain with Rapamycin FRB ectodomain 2 aa 
            scaffold, 6 aa linker domain, C-terminal TEV protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6328)
  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 6328)
  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: FRB_SCF2_LD6_CTEV
FEATURES             Location/Qualifiers
     source          1..6328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     polyA_signal    1604..1996
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      2904..3417
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2991..3010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3201..3222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3898..3917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4017..4039
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4077..4095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4163..4267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4268..4336,4337..5128)
                     /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(4486..4505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5299..5887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5788..5807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6127..6256
                     /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 gggatgctcg agatgtggca tgaaggcctg
      781 gaagaggcat ctcgtttgta ctttggggaa aggaacgtga aaggcatgtt tgaggtgctg
      841 gagcccttgc atgctatgat ggaacggggc ccccagactc tgaaggaaac atcctttaat
      901 caggcctatg gtcgagattt aatggaggcc caagagtggt gcaggaagta catgaaatca
      961 gggaatgtca aggacctcct ccaagcctgg gacctctatt atcatgtgtt ccgacgaatc
     1021 tcaaagacta gaaccggtaa gcttttttgg gcactggtcg tggttgctgg agtcctgttt
     1081 tgttatggct tgctagtgac agtggctctt tgtgttatct gggtaagatc tggtatgtct
     1141 agcatggtgt cagacactag ttgcacattc ccttcatctg atggcatatt ctggaagcat
     1201 tggattcaaa ccaaggatgg gcagtgtggc agtccattag tatcaactag agatgggttc
     1261 attgttggta tacactcagc atcgaatttc accaacacaa acaattattt cacaagcgtg
     1321 ccgaaaaact tcatggaatt gttgacaaat caggaggcgc agcagtgggt tagtggttgg
     1381 cgattaaatg ctgactcagt attgtggggg ggccataaag ttttcatggt gaaacctgaa
     1441 gagccttttc agccagttaa ggaagcgact caactcatga attaggcggc cgccaccgcg
     1501 gtggagctgt ttaaacctcg agcaattcac cccaccagtg caggctgcct atcagaaagt
     1561 ggtggctggt gtggctaatg ccctggccca caagtatcac taagctcgct ttcttgctgt
     1621 ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg gggatattat
     1681 gaagggcctt gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat
     1741 gtatttaaat tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta
     1801 aaacataaag aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc
     1861 catgaaagaa ggtgaggctg caaacagtaa tgcacattgg caacagcccc tgatgcctat
     1921 gccttattca tccctcagaa aaggattcaa gtagaggctt gatttggagg ttaaagtttt
     1981 gctatgctgt attttacatt acttattgtt ttagctgtcc tcatgaatgt cttttcacta
     2041 cccatttgct tatcctgcat ctctcagcct tgactccact cagttctctt gcttagagat
     2101 accacctttc ccctgaagtg ttccttccat gttttacggc gagatggttt ctcctcgcct
     2161 ggccactcag ccttagttgt ctctgttgtc ttatagaggt ctacttgaag aaggaaaaac
     2221 agggggcatg gtttgactgt cctgtgagcc cttcttccct gcctccccca ctcacagtga
     2281 cccggaatcc ctcgacatgg cagtctagag catggctacg tagataagta gcatggcggg
     2341 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2401 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2461 gcctcagtga gcgagcgagc gcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2521 cccttcccaa cagttgcgca gcctgaatgg cgaatggaat tccagacgat tgagcgtcaa
     2581 aatgtaggta tttccatgag cgtttttcct gttgcaatgg ctggcggtaa tattgttctg
     2641 gatattacca gcaaggccga tagtttgagt tcttctactc aggcaagtga tgttattact
     2701 aatcaaagaa gtattgcgac aacggttaat ttgcgtgatg gacagactct tttactcggt
     2761 ggcctcactg attataaaaa cacttctcag gattctggcg taccgttcct gtctaaaatc
     2821 cctttaatcg gcctcctgtt tagctcccgc tctgattcta acgaggaaag cacgttatac
     2881 gtgctcgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2941 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3001 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3061 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3121 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3181 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3241 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3301 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta
     3361 aatatttgct tatacaatct tcctgttttt ggggcttttc tgattatcaa ccggggtaca
     3421 tatgattgac atgctagttt tacgattacc gttcatcgat tctcttgttt gctccagact
     3481 ctcaggcaat gacctgatag cctttgtaga gacctctcaa aaatagctac cctctccggc
     3541 atgaatttat cagctagaac ggttgaatat catattgatg gtgatttgac tgtctccggc
     3601 ctttctcacc cgtttgaatc tttacctaca cattactcag gcattgcatt taaaatatat
     3661 gagggttcta aaaattttta tccttgcgtt gaaataaagg cttctcccgc aaaagtatta
     3721 cagggtcata atgtttttgg tacaaccgat ttagctttat gctctgaggc tttattgctt
     3781 aattttgcta attctttgcc ttgcctgtat gatttattgg atgttggaat tcctgatgcg
     3841 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     3901 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3961 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4021 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     4081 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4141 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4201 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4261 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4321 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4381 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4441 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4501 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4561 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4621 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4681 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4741 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4801 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4861 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4921 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4981 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5041 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5101 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5161 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5221 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5281 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5341 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5401 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5461 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5521 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5581 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5641 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5701 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5761 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5821 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5881 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5941 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6001 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6061 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6121 cagcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
     6181 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     6241 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
     6301 tgctctaggg atccggccag atatacgc
//
