LOCUS       sequence-161519-        7194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA protease chain with V1-MESA ectodomain, 45 extracellular 
            linkers, a flag tag, and Tev protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7194)
  AUTHORS   Schwarz KA, Daringer NM, Dolberg TB, Leonard JN
  TITLE     Rewiring human cellular input-output using modular extracellular 
            sensors.
  JOURNAL   Nat Chem Biol. 2016 Dec 12. doi: 10.1038/nchembio.2253.
  PUBMED    27941759
REFERENCE   2  (bases 1 to 7194)
  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: "Nat Chem 
            Biol. 2016 Dec 12. doi: 10.1038/nchembio.2253."
COMMENT     Sequence Label: V1-MESA-45F-Tev
FEATURES             Location/Qualifiers
     source          1..7194
                     /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             1813..2517
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     polyA_signal    2641..3033
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      3941..4454
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4238..4259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4935..4954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5054..5076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5114..5132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5200..5304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5305..5373,5374..6165)
                     /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(5523..5542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6336..6924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6825..6844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7164..7194,1..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 gccatcatgg cttggaccag cttaattctt agcttgcttg ccctgtgcag cggtgcatca
      901 tctgacatac agatgactca gtcacctagc agtctgtctg cttcagtcgg tgatagggta
      961 actatcacct gtcgggcatc tcaggatgtg tccacggcag ttgcctggta ccagcagaag
     1021 cccggcaagg cgcctaagtt gcttatctat tcagcttcat ttttgtatag tggggtgcct
     1081 agccggttta gcggaagtgg ttctggcaca gacttcactt tgacaatcag ttccctccaa
     1141 ccggaggatt tcgctaccta ctactgtcag caggggtacg gaaacccctt tacgtttgga
     1201 cagggcacta aagtggagat taagagggga ggcggaggct ccggtggggg aggctctggt
     1261 ggtggaggta gcgaggttca gctggtggaa agtggtggcg gactagtaca gccaggcggc
     1321 tccctgcggc tgagttgcgc cgcgagtggc tttactatat ctgactattg gatccattgg
     1381 gtcaggcaag cccctgggaa aggattggag tgggttgctg gtattacccc cgcaggtggg
     1441 tatacttatt atgcagatag cgtgaaaggg aggttcacca tctctgcaga cacaagcaag
     1501 aacacggcct acctacagat gaactctctg agggcagagg acactgccgt ctactattgc
     1561 gcccgatttg tgttttttct accctacgcc atggactatt ggggacaggg gacattggta
     1621 ggcgccggga caggagggtc tgggggtggc actggcggat ctggaggaga ctacaaggac
     1681 gatgacgaca acggcggatc gggggggtct ggtggtagtg gcggttcggg gggaggtacc
     1741 ggtctggtcg tggttgctgg agtcctgttt tgttatggct tgctagtgac agtggctctt
     1801 tgtgttgaaa acgagagctt gtttaagggg ccgcgtgatt acaacccgat atcgagcacc
     1861 atttgtcatt tgacgaatga atctgatggg cacacaacat cgttgtatgg tattggattt
     1921 ggtcccttca tcattacaaa caagcacttg tttagaagaa ataatggaac actgttggtc
     1981 caatcactac atggtgtatt caaggtcaag aacaccacga ctttgcaaca acacctcatt
     2041 gatgggaggg acatgataat tattcgcatg cctaaggatt tcccaccatt tcctcaaaag
     2101 ctgaaattta gagagccaca aagggaagag cgcatatgtc ttgtgacaac caacttccaa
     2161 actaagagca tgtctagcat ggtgtcagac actagttgca cattcccttc atctgatggc
     2221 atattctgga agcattggat tcaaaccaag gatgggcagt gtggcagtcc attagtatca
     2281 actagagatg ggttcattgt tggtatacac tcagcatcga atttcaccaa cacaaacaat
     2341 tatttcacaa gcgtgccgaa aaacttcatg gaattgttga caaatcagga ggcgcagcag
     2401 tgggttagtg gttggcgatt aaatgctgac tcagtattgt gggggggcca taaagttttc
     2461 atggtgaaac ctgaagagcc ttttcagcca gttaaggaag cgactcaact catgaattag
     2521 tagcggccgc caccgcggtg gagctgttta aacctcgagc aattcacccc accagtgcag
     2581 gctgcctatc agaaagtggt ggctggtgtg gctaatgccc tggcccacaa gtatcactaa
     2641 gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac
     2701 taaactgggg atattatgaa gggccttgag catctggatt ctgcctaata aaaaacattt
     2761 attttcattg caatgatgta tttaaattat ttctgaatat tttactaaaa agggaatgtg
     2821 ggaggtcagt gcatttaaaa cataaagaaa tgaagagcta gttcaaacct tgggaaaata
     2881 cactatatct taaactccat gaaagaaggt gaggctgcaa acagtaatgc acattggcaa
     2941 cagcccctga tgcctatgcc ttattcatcc ctcagaaaag gattcaagta gaggcttgat
     3001 ttggaggtta aagttttgct atgctgtatt ttacattact tattgtttta gctgtcctca
     3061 tgaatgtctt ttcactaccc atttgcttat cctgcatctc tcagccttga ctccactcag
     3121 ttctcttgct tagagatacc acctttcccc tgaagtgttc cttccatgtt ttacggcgag
     3181 atggtttctc ctcgcctggc cactcagcct tagttgtctc tgttgtctta tagaggtcta
     3241 cttgaagaag gaaaaacagg gggcatggtt tgactgtcct gtgagccctt cttccctgcc
     3301 tcccccactc acagtgaccc ggaatccctc gacatggcag tctagagcat ggctacgtag
     3361 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3421 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3481 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg ccagctggcg taatagcgaa
     3541 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaattcc
     3601 agacgattga gcgtcaaaat gtaggtattt ccatgagcgt ttttcctgtt gcaatggctg
     3661 gcggtaatat tgttctggat attaccagca aggccgatag tttgagttct tctactcagg
     3721 caagtgatgt tattactaat caaagaagta ttgcgacaac ggttaatttg cgtgatggac
     3781 agactctttt actcggtggc ctcactgatt ataaaaacac ttctcaggat tctggcgtac
     3841 cgttcctgtc taaaatccct ttaatcggcc tcctgtttag ctcccgctct gattctaacg
     3901 aggaaagcac gttatacgtg ctcgtcaaag caaccatagt acgcgccctg tagcggcgca
     3961 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4021 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4081 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4141 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4201 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4261 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4321 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4381 ttaacgttta caatttaaat atttgcttat acaatcttcc tgtttttggg gcttttctga
     4441 ttatcaaccg gggtacatat gattgacatg ctagttttac gattaccgtt catcgattct
     4501 cttgtttgct ccagactctc aggcaatgac ctgatagcct ttgtagagac ctctcaaaaa
     4561 tagctaccct ctccggcatg aatttatcag ctagaacggt tgaatatcat attgatggtg
     4621 atttgactgt ctccggcctt tctcacccgt ttgaatcttt acctacacat tactcaggca
     4681 ttgcatttaa aatatatgag ggttctaaaa atttttatcc ttgcgttgaa ataaaggctt
     4741 ctcccgcaaa agtattacag ggtcataatg tttttggtac aaccgattta gctttatgct
     4801 ctgaggcttt attgcttaat tttgctaatt ctttgccttg cctgtatgat ttattggatg
     4861 ttggaattcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4921 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4981 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5041 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5101 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5161 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5221 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5281 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5341 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5401 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5461 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5521 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5581 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5641 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5701 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5761 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5821 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5881 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5941 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6001 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6061 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6121 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6181 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6241 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6301 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6361 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6421 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6481 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6541 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6601 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6661 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6721 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6781 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6841 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6901 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6961 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7021 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7081 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7141 tggccgattc attaatgcag cagctgcgcg ctcgctcgct cactgaggcc gccc
//
