LOCUS       sequence-138321-        7081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA protease chain with dTomato ectodomain and no linker domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  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 7081)
  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: dTomato_LD 0_TEV
FEATURES             Location/Qualifiers
     source          1..7081
                     /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..1623
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(1055..1073)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1323..1342
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1517..1540
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1702..2406
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     polyA_signal    2528..2920
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      3828..4341
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3915..3934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4125..4146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4822..4841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4941..4963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5001..5019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5087..5191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5192..5260,5261..6052)
                     /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(5410..5429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6223..6811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6712..6731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7051..7081,1..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 aggggtgagc aagggcgagg aggtcatcaa agagttcatg
      961 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     1021 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     1081 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     1141 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     1201 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     1261 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     1321 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     1381 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     1441 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     1501 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     1561 cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac
     1621 aagaagcttt tttgggcact ggtcgtggtt gctggagtcc tgttttgtta tggcttgcta
     1681 gtgacagtgg ctctttgtgt tgagagcttg tttaaggggc cgcgtgatta caacccgata
     1741 tcgagcacca tttgtcattt gacgaatgaa tctgatgggc acacaacatc gttgtatggt
     1801 attggatttg gtcccttcat cattacaaac aagcacttgt ttagaagaaa taatggaaca
     1861 ctgttggtcc aatcactaca tggtgtattc aaggtcaaga acaccacgac tttgcaacaa
     1921 cacctcattg atgggaggga catgataatt attcgcatgc ctaaggattt cccaccattt
     1981 cctcaaaagc tgaaatttag agagccacaa agggaagagc gcatatgtct tgtgacaacc
     2041 aacttccaaa ctaagagcat gtctagcatg gtgtcagaca ctagttgcac attcccttca
     2101 tctgatggca tattctggaa gcattggatt caaaccaagg atgggcagtg tggcagtcca
     2161 ttagtatcaa ctagagatgg gttcattgtt ggtatacact cagcatcgaa tttcaccaac
     2221 acaaacaatt atttcacaag cgtgccgaaa aacttcatgg aattgttgac aaatcaggag
     2281 gcgcagcagt gggttagtgg ttggcgatta aatgctgact cagtattgtg ggggggccat
     2341 aaagttttca tggtgaaacc tgaagagcct tttcagccag ttaaggaagc gactcaactc
     2401 atgaattagg cggccgccac cgcggtggag ctgtttaaac ctcgagcaat tcaccccacc
     2461 agtgcaggct gcctatcaga aagtggtggc tggtgtggct aatgccctgg cccacaagta
     2521 tcactaagct cgctttcttg ctgtccaatt tctattaaag gttcctttgt tccctaagtc
     2581 caactactaa actggggata ttatgaaggg ccttgagcat ctggattctg cctaataaaa
     2641 aacatttatt ttcattgcaa tgatgtattt aaattatttc tgaatatttt actaaaaagg
     2701 gaatgtggga ggtcagtgca tttaaaacat aaagaaatga agagctagtt caaaccttgg
     2761 gaaaatacac tatatcttaa actccatgaa agaaggtgag gctgcaaaca gtaatgcaca
     2821 ttggcaacag cccctgatgc ctatgcctta ttcatccctc agaaaaggat tcaagtagag
     2881 gcttgatttg gaggttaaag ttttgctatg ctgtatttta cattacttat tgttttagct
     2941 gtcctcatga atgtcttttc actacccatt tgcttatcct gcatctctca gccttgactc
     3001 cactcagttc tcttgcttag agataccacc tttcccctga agtgttcctt ccatgtttta
     3061 cggcgagatg gtttctcctc gcctggccac tcagccttag ttgtctctgt tgtcttatag
     3121 aggtctactt gaagaaggaa aaacaggggg catggtttga ctgtcctgtg agcccttctt
     3181 ccctgcctcc cccactcaca gtgacccgga atccctcgac atggcagtct agagcatggc
     3241 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3301 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3361 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcca gctggcgtaa
     3421 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3481 gaattccaga cgattgagcg tcaaaatgta ggtatttcca tgagcgtttt tcctgttgca
     3541 atggctggcg gtaatattgt tctggatatt accagcaagg ccgatagttt gagttcttct
     3601 actcaggcaa gtgatgttat tactaatcaa agaagtattg cgacaacggt taatttgcgt
     3661 gatggacaga ctcttttact cggtggcctc actgattata aaaacacttc tcaggattct
     3721 ggcgtaccgt tcctgtctaa aatcccttta atcggcctcc tgtttagctc ccgctctgat
     3781 tctaacgagg aaagcacgtt atacgtgctc gtcaaagcaa ccatagtacg cgccctgtag
     3841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3901 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4021 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4141 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4201 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4261 caaaatatta acgtttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct
     4321 tttctgatta tcaaccgggg tacatatgat tgacatgcta gttttacgat taccgttcat
     4381 cgattctctt gtttgctcca gactctcagg caatgacctg atagcctttg tagagacctc
     4441 tcaaaaatag ctaccctctc cggcatgaat ttatcagcta gaacggttga atatcatatt
     4501 gatggtgatt tgactgtctc cggcctttct cacccgtttg aatctttacc tacacattac
     4561 tcaggcattg catttaaaat atatgagggt tctaaaaatt tttatccttg cgttgaaata
     4621 aaggcttctc ccgcaaaagt attacagggt cataatgttt ttggtacaac cgatttagct
     4681 ttatgctctg aggctttatt gcttaatttt gctaattctt tgccttgcct gtatgattta
     4741 ttggatgttg gaattcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4801 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4861 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4921 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4981 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5041 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5401 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5701 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6361 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6961 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7021 cgcgcgttgg ccgattcatt aatgcagcag ctgcgcgctc gctcgctcac tgaggccgcc
     7081 c
//
