LOCUS       sequence-94236-e        8151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA target chain with dTomato ectodomain, no linker domain, M 
            cleavage sequence, and tTA-BFP fusion .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8151)
  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 8151)
  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_CS M_tTA-BFP
FEATURES             Location/Qualifiers
     source          1..8151
                     /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..1722
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             1720..2343
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             join(1723..2343,2344..2727)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(1796..1815)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             join(2758..2760,2761..2763,2764..3477)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(2803..2824)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3598..3990
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      4898..5411
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4985..5004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5195..5216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5892..5911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6011..6033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6071..6089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6157..6261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6262..6330,6331..7122)
                     /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(6480..6499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7293..7881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7782..7801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(8121..8151,1..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 tgaaaacctg tattttcaga tgatgtctag attagataaa
     1741 agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga aggtttaaca
     1801 acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa
     1861 aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca ccatactcac
     1921 ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt
     1981 agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca
     2041 gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca
     2101 ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg ttgcgtattg
     2161 gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg
     2221 ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga gccagccttc
     2281 ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtggg
     2341 tccgcgtaca gccgcgcgcg tacgaaaaac aattacgggt ctaccatcga gggcctgctc
     2401 gatctcccgg acgacgacgc ccccgaagag gcggggctgg cggctccgcg cctgtccttt
     2461 ctccccgcgg gacacacgcg cagactgtcg acggcccccc cgaccgatgt cagcctgggg
     2521 gacgagctcc acttagacgg cgaggacgtg gcgatggcgc atgccgacgc gctagacgat
     2581 ttcgatctgg acatgttggg ggacggggat tccccgggtc cgggatttac cccccacgac
     2641 tccgccccct acggcgctct ggatatggcc gacttcgagt ttgagcagat gtttaccgat
     2701 gcccttggaa ttgacgagta cggtggaggt accggcggag gctccggtgg tggctctatg
     2761 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2821 gacgtaaacg gccacaagtt cagcgtgagg ggcgagggcg agggcgatgc caccaacggc
     2881 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2941 gtgaccaccc tgagccacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     3001 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3061 aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctagtg
     3121 aaccgcatcg agctgaaggg cgtcgacttc aaggaggacg gcaacatcct ggggcacaag
     3181 ctggagtaca acttcaacag ccacaacatc tatatcatgg ccgtcaagca gaagaacggc
     3241 atcaaggtga acttcaagat ccgccacaac gtggaggacg gcagcgtgca gctcgccgac
     3301 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga cagccactac
     3361 ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3421 ctggagttcc gcaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatag
     3481 cggccgccac cgcggtggag ctgtttaaac ctcgagcaat tcaccccacc agtgcaggct
     3541 gcctatcaga aagtggtggc tggtgtggct aatgccctgg cccacaagta tcactaagct
     3601 cgctttcttg ctgtccaatt tctattaaag gttcctttgt tccctaagtc caactactaa
     3661 actggggata ttatgaaggg ccttgagcat ctggattctg cctaataaaa aacatttatt
     3721 ttcattgcaa tgatgtattt aaattatttc tgaatatttt actaaaaagg gaatgtggga
     3781 ggtcagtgca tttaaaacat aaagaaatga agagctagtt caaaccttgg gaaaatacac
     3841 tatatcttaa actccatgaa agaaggtgag gctgcaaaca gtaatgcaca ttggcaacag
     3901 cccctgatgc ctatgcctta ttcatccctc agaaaaggat tcaagtagag gcttgatttg
     3961 gaggttaaag ttttgctatg ctgtatttta cattacttat tgttttagct gtcctcatga
     4021 atgtcttttc actacccatt tgcttatcct gcatctctca gccttgactc cactcagttc
     4081 tcttgcttag agataccacc tttcccctga agtgttcctt ccatgtttta cggcgagatg
     4141 gtttctcctc gcctggccac tcagccttag ttgtctctgt tgtcttatag aggtctactt
     4201 gaagaaggaa aaacaggggg catggtttga ctgtcctgtg agcccttctt ccctgcctcc
     4261 cccactcaca gtgacccgga atccctcgac atggcagtct agagcatggc tacgtagata
     4321 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     4381 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4441 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcca gctggcgtaa tagcgaagag
     4501 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaattccaga
     4561 cgattgagcg tcaaaatgta ggtatttcca tgagcgtttt tcctgttgca atggctggcg
     4621 gtaatattgt tctggatatt accagcaagg ccgatagttt gagttcttct actcaggcaa
     4681 gtgatgttat tactaatcaa agaagtattg cgacaacggt taatttgcgt gatggacaga
     4741 ctcttttact cggtggcctc actgattata aaaacacttc tcaggattct ggcgtaccgt
     4801 tcctgtctaa aatcccttta atcggcctcc tgtttagctc ccgctctgat tctaacgagg
     4861 aaagcacgtt atacgtgctc gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4921 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4981 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5041 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5101 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5161 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5221 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5281 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5341 acgtttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct tttctgatta
     5401 tcaaccgggg tacatatgat tgacatgcta gttttacgat taccgttcat cgattctctt
     5461 gtttgctcca gactctcagg caatgacctg atagcctttg tagagacctc tcaaaaatag
     5521 ctaccctctc cggcatgaat ttatcagcta gaacggttga atatcatatt gatggtgatt
     5581 tgactgtctc cggcctttct cacccgtttg aatctttacc tacacattac tcaggcattg
     5641 catttaaaat atatgagggt tctaaaaatt tttatccttg cgttgaaata aaggcttctc
     5701 ccgcaaaagt attacagggt cataatgttt ttggtacaac cgatttagct ttatgctctg
     5761 aggctttatt gcttaatttt gctaattctt tgccttgcct gtatgattta ttggatgttg
     5821 gaattcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     5881 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5941 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6001 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6061 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6121 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6181 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6241 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6301 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6361 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6421 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6481 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6541 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6601 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6661 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6721 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6781 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6841 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6901 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6961 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7021 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7081 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7141 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7201 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7261 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7381 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     7441 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7501 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7561 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7621 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7681 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7741 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7801 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7861 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7921 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7981 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8041 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8101 ccgattcatt aatgcagcag ctgcgcgctc gctcgctcac tgaggccgcc c
//
