LOCUS       Exported                7167 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  MESA protease chain with V2-MESA ectodomain, 35 extracellular 
            linkers, a flag tag, and Tev protease.
ACCESSION   .
VERSION     .
KEYWORDS    V2-MESA-35F-Tev
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7167)
  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 7167)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7167
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1786..2490
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     polyA_signal    2614..3006
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      3914..4427
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4001..4020)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4211..4232
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4908..4927)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5027..5049
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5087..5105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5173..5277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5278..6138
                     /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(5496..5515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6309..6897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6798..6817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7051..7068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   7137..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 tcaggtgatc agaaggtctc tggcctggta tcagcagaag
     1021 cccggcaagg ctcctaagtt gcttatctat gcagcttcaa acttggccag tggggtgcct
     1081 agccggttta gcggaagtgg ttctggcaca gacttcactt tgacaatcag ttccctccaa
     1141 ccggaggatt tcgctaccta ctactgtcag cagtccaaca ccagccctct gacgtttgga
     1201 cagggcacta aagtggagat taagagggga ggcggaggct ccggtggggg aggctctggt
     1261 ggtggaggta gcgaggttca gctggtggaa agtggtggcg gactagtaca gccaggcggc
     1321 tccctgcggc tgagttgcgc cgcgagtggc ttttctataa acggatcctg gatcttctgg
     1381 gtcaggcaag cccctgggaa aggattggag tgggttggag ccatttggcc tttcggtggg
     1441 tatactcact atgcagatag cgtgaaaggg aggttcacca tctctgcaga cacaagcaag
     1501 aacacggcct acctacagat gaactctctg agggcagagg acactgccgt ctactattgc
     1561 gcccgatggg gacactccac aagcccttgg gccatggact attggggaca ggggacattg
     1621 gtaggcgccg gatctggagg agactacaag gacgatgacg acaacggcgg atcggggggg
     1681 tctggtggta gtggcggttc ggggggaggt accggtctgg tcgtggttgc tggagtcctg
     1741 ttttgttatg gcttgctagt gacagtggct ctttgtgttg aaaacgagag cttgtttaag
     1801 gggccgcgtg attacaaccc gatatcgagc accatttgtc atttgacgaa tgaatctgat
     1861 gggcacacaa catcgttgta tggtattgga tttggtccct tcatcattac aaacaagcac
     1921 ttgtttagaa gaaataatgg aacactgttg gtccaatcac tacatggtgt attcaaggtc
     1981 aagaacacca cgactttgca acaacacctc attgatggga gggacatgat aattattcgc
     2041 atgcctaagg atttcccacc atttcctcaa aagctgaaat ttagagagcc acaaagggaa
     2101 gagcgcatat gtcttgtgac aaccaacttc caaactaaga gcatgtctag catggtgtca
     2161 gacactagtt gcacattccc ttcatctgat ggcatattct ggaagcattg gattcaaacc
     2221 aaggatgggc agtgtggcag tccattagta tcaactagag atgggttcat tgttggtata
     2281 cactcagcat cgaatttcac caacacaaac aattatttca caagcgtgcc gaaaaacttc
     2341 atggaattgt tgacaaatca ggaggcgcag cagtgggtta gtggttggcg attaaatgct
     2401 gactcagtat tgtggggggg ccataaagtt ttcatggtga aacctgaaga gccttttcag
     2461 ccagttaagg aagcgactca actcatgaat tagtagcggc cgccaccgcg gtggagctgt
     2521 ttaaacctcg agcaattcac cccaccagtg caggctgcct atcagaaagt ggtggctggt
     2581 gtggctaatg ccctggccca caagtatcac taagctcgct ttcttgctgt ccaatttcta
     2641 ttaaaggttc ctttgttccc taagtccaac tactaaactg gggatattat gaagggcctt
     2701 gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat gtatttaaat
     2761 tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta aaacataaag
     2821 aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc catgaaagaa
     2881 ggtgaggctg caaacagtaa tgcacattgg caacagcccc tgatgcctat gccttattca
     2941 tccctcagaa aaggattcaa gtagaggctt gatttggagg ttaaagtttt gctatgctgt
     3001 attttacatt acttattgtt ttagctgtcc tcatgaatgt cttttcacta cccatttgct
     3061 tatcctgcat ctctcagcct tgactccact cagttctctt gcttagagat accacctttc
     3121 ccctgaagtg ttccttccat gttttacggc gagatggttt ctcctcgcct ggccactcag
     3181 ccttagttgt ctctgttgtc ttatagaggt ctacttgaag aaggaaaaac agggggcatg
     3241 gtttgactgt cctgtgagcc cttcttccct gcctccccca ctcacagtga cccggaatcc
     3301 ctcgacatgg cagtctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3361 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3421 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3481 gcgagcgagc gcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3541 cagttgcgca gcctgaatgg cgaatggaat tccagacgat tgagcgtcaa aatgtaggta
     3601 tttccatgag cgtttttcct gttgcaatgg ctggcggtaa tattgttctg gatattacca
     3661 gcaaggccga tagtttgagt tcttctactc aggcaagtga tgttattact aatcaaagaa
     3721 gtattgcgac aacggttaat ttgcgtgatg gacagactct tttactcggt ggcctcactg
     3781 attataaaaa cacttctcag gattctggcg taccgttcct gtctaaaatc cctttaatcg
     3841 gcctcctgtt tagctcccgc tctgattcta acgaggaaag cacgttatac gtgctcgtca
     3901 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3961 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4021 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4081 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4141 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4201 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4261 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4321 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aatatttgct
     4381 tatacaatct tcctgttttt ggggcttttc tgattatcaa ccggggtaca tatgattgac
     4441 atgctagttt tacgattacc gttcatcgat tctcttgttt gctccagact ctcaggcaat
     4501 gacctgatag cctttgtaga gacctctcaa aaatagctac cctctccggc atgaatttat
     4561 cagctagaac ggttgaatat catattgatg gtgatttgac tgtctccggc ctttctcacc
     4621 cgtttgaatc tttacctaca cattactcag gcattgcatt taaaatatat gagggttcta
     4681 aaaattttta tccttgcgtt gaaataaagg cttctcccgc aaaagtatta cagggtcata
     4741 atgtttttgg tacaaccgat ttagctttat gctctgaggc tttattgctt aattttgcta
     4801 attctttgcc ttgcctgtat gatttattgg atgttggaat tcctgatgcg gtattttctc
     4861 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4921 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4981 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5041 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5101 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5161 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5221 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5281 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5341 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5401 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5461 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5521 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5581 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5641 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5701 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5761 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5821 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5881 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5941 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6001 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6061 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6121 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6421 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7021 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7081 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcagctgc
     7141 gcgctcgctc gctcactgag gccgccc
//
