LOCUS       sequence-94235-e        8160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA target chain with mCherry 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 8160)
  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 8160)
  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: mCherry_LD 0_CS M_tTA-BFP
FEATURES             Location/Qualifiers
     source          1..8160
                     /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..1632
                     /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(1070..1088)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1338..1357
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1711..1731
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             1729..2352
                     /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(1732..2352,2353..2736)
                     /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(1805..1824)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             join(2767..2769,2770..2772,2773..3486)
                     /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(2812..2833)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3607..3999
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      4907..5420
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4994..5013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5204..5225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5901..5920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6020..6042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6080..6098)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6166..6270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6271..6339,6340..7131)
                     /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(6489..6508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7302..7890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7791..7810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(8130..8160,1..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 agggatggtg agcaagggcg aggaggataa catggccatc
      961 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     1021 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1081 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     1141 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     1201 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1261 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1321 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1381 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1441 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1501 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     1561 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1621 gagctgtaca agaagctttt ttgggcactg gtcgtggttg ctggagtcct gttttgttat
     1681 ggcttgctag tgacagtggc tctttgtgtt gaaaacctgt attttcagat gatgtctaga
     1741 ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt cggaatcgaa
     1801 ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac attgtattgg
     1861 catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt agataggcac
     1921 catactcact tttgcccttt agaaggggaa agctggcaag attttttacg taataacgct
     1981 aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca tttaggtaca
     2041 cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt atgccaacaa
     2101 ggtttttcac tagagaatgc attatatgca ctcagcgctg tggggcattt tactttaggt
     2161 tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac acctactact
     2221 gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca aggtgcagag
     2281 ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca acttaaatgt
     2341 gaaagtgggt ccgcgtacag ccgcgcgcgt acgaaaaaca attacgggtc taccatcgag
     2401 ggcctgctcg atctcccgga cgacgacgcc cccgaagagg cggggctggc ggctccgcgc
     2461 ctgtcctttc tccccgcggg acacacgcgc agactgtcga cggccccccc gaccgatgtc
     2521 agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg
     2581 ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc
     2641 ccccacgact ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg
     2701 tttaccgatg cccttggaat tgacgagtac ggtggaggta ccggcggagg ctccggtggt
     2761 ggctctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2821 ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc
     2881 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2941 cccaccctcg tgaccaccct gagccacggc gtgcagtgct tcgcccgcta ccccgaccac
     3001 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3061 atcttcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     3121 accctagtga accgcatcga gctgaagggc gtcgacttca aggaggacgg caacatcctg
     3181 gggcacaagc tggagtacaa cttcaacagc cacaacatct atatcatggc cgtcaagcag
     3241 aagaacggca tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag
     3301 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3361 agccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac
     3421 atggtcctgc tggagttccg caccgccgcc gggatcactc tcggcatgga cgagctgtac
     3481 aagtaatagc ggccgccacc gcggtggagc tgtttaaacc tcgagcaatt caccccacca
     3541 gtgcaggctg cctatcagaa agtggtggct ggtgtggcta atgccctggc ccacaagtat
     3601 cactaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt ccctaagtcc
     3661 aactactaaa ctggggatat tatgaagggc cttgagcatc tggattctgc ctaataaaaa
     3721 acatttattt tcattgcaat gatgtattta aattatttct gaatatttta ctaaaaaggg
     3781 aatgtgggag gtcagtgcat ttaaaacata aagaaatgaa gagctagttc aaaccttggg
     3841 aaaatacact atatcttaaa ctccatgaaa gaaggtgagg ctgcaaacag taatgcacat
     3901 tggcaacagc ccctgatgcc tatgccttat tcatccctca gaaaaggatt caagtagagg
     3961 cttgatttgg aggttaaagt tttgctatgc tgtattttac attacttatt gttttagctg
     4021 tcctcatgaa tgtcttttca ctacccattt gcttatcctg catctctcag ccttgactcc
     4081 actcagttct cttgcttaga gataccacct ttcccctgaa gtgttccttc catgttttac
     4141 ggcgagatgg tttctcctcg cctggccact cagccttagt tgtctctgtt gtcttataga
     4201 ggtctacttg aagaaggaaa aacagggggc atggtttgac tgtcctgtga gcccttcttc
     4261 cctgcctccc ccactcacag tgacccggaa tccctcgaca tggcagtcta gagcatggct
     4321 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     4381 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4441 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgccag ctggcgtaat
     4501 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4561 aattccagac gattgagcgt caaaatgtag gtatttccat gagcgttttt cctgttgcaa
     4621 tggctggcgg taatattgtt ctggatatta ccagcaaggc cgatagtttg agttcttcta
     4681 ctcaggcaag tgatgttatt actaatcaaa gaagtattgc gacaacggtt aatttgcgtg
     4741 atggacagac tcttttactc ggtggcctca ctgattataa aaacacttct caggattctg
     4801 gcgtaccgtt cctgtctaaa atccctttaa tcggcctcct gtttagctcc cgctctgatt
     4861 ctaacgagga aagcacgtta tacgtgctcg tcaaagcaac catagtacgc gccctgtagc
     4921 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4981 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5041 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5101 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5161 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5221 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5281 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5341 aaaatattaa cgtttacaat ttaaatattt gcttatacaa tcttcctgtt tttggggctt
     5401 ttctgattat caaccggggt acatatgatt gacatgctag ttttacgatt accgttcatc
     5461 gattctcttg tttgctccag actctcaggc aatgacctga tagcctttgt agagacctct
     5521 caaaaatagc taccctctcc ggcatgaatt tatcagctag aacggttgaa tatcatattg
     5581 atggtgattt gactgtctcc ggcctttctc acccgtttga atctttacct acacattact
     5641 caggcattgc atttaaaata tatgagggtt ctaaaaattt ttatccttgc gttgaaataa
     5701 aggcttctcc cgcaaaagta ttacagggtc ataatgtttt tggtacaacc gatttagctt
     5761 tatgctctga ggctttattg cttaattttg ctaattcttt gccttgcctg tatgatttat
     5821 tggatgttgg aattcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5881 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5941 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6001 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6061 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6121 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6241 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6301 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6361 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6421 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6481 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6541 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6601 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6661 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6721 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6781 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6841 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6901 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6961 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7021 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7441 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7921 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7981 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8041 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8101 gcgcgttggc cgattcatta atgcagcagc tgcgcgctcg ctcgctcact gaggccgccc
//
