LOCUS       Exported                7090 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  MESA protease chain with mCherry ectodomain and no linker domain.
ACCESSION   .
VERSION     .
KEYWORDS    mCherry_LD 0_TEV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7090)
  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 7090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7090
                     /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             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..2415
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     polyA_signal    2537..2929
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      3837..4350
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3924..3943)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4134..4155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4831..4850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4950..4972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5010..5028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5096..5200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5201..6061
                     /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(5419..5438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6232..6820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6721..6740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6974..6991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   7060..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 gagagcttgt ttaaggggcc gcgtgattac
     1741 aacccgatat cgagcaccat ttgtcatttg acgaatgaat ctgatgggca cacaacatcg
     1801 ttgtatggta ttggatttgg tcccttcatc attacaaaca agcacttgtt tagaagaaat
     1861 aatggaacac tgttggtcca atcactacat ggtgtattca aggtcaagaa caccacgact
     1921 ttgcaacaac acctcattga tgggagggac atgataatta ttcgcatgcc taaggatttc
     1981 ccaccatttc ctcaaaagct gaaatttaga gagccacaaa gggaagagcg catatgtctt
     2041 gtgacaacca acttccaaac taagagcatg tctagcatgg tgtcagacac tagttgcaca
     2101 ttcccttcat ctgatggcat attctggaag cattggattc aaaccaagga tgggcagtgt
     2161 ggcagtccat tagtatcaac tagagatggg ttcattgttg gtatacactc agcatcgaat
     2221 ttcaccaaca caaacaatta tttcacaagc gtgccgaaaa acttcatgga attgttgaca
     2281 aatcaggagg cgcagcagtg ggttagtggt tggcgattaa atgctgactc agtattgtgg
     2341 gggggccata aagttttcat ggtgaaacct gaagagcctt ttcagccagt taaggaagcg
     2401 actcaactca tgaattaggc ggccgccacc gcggtggagc tgtttaaacc tcgagcaatt
     2461 caccccacca gtgcaggctg cctatcagaa agtggtggct ggtgtggcta atgccctggc
     2521 ccacaagtat cactaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt
     2581 ccctaagtcc aactactaaa ctggggatat tatgaagggc cttgagcatc tggattctgc
     2641 ctaataaaaa acatttattt tcattgcaat gatgtattta aattatttct gaatatttta
     2701 ctaaaaaggg aatgtgggag gtcagtgcat ttaaaacata aagaaatgaa gagctagttc
     2761 aaaccttggg aaaatacact atatcttaaa ctccatgaaa gaaggtgagg ctgcaaacag
     2821 taatgcacat tggcaacagc ccctgatgcc tatgccttat tcatccctca gaaaaggatt
     2881 caagtagagg cttgatttgg aggttaaagt tttgctatgc tgtattttac attacttatt
     2941 gttttagctg tcctcatgaa tgtcttttca ctacccattt gcttatcctg catctctcag
     3001 ccttgactcc actcagttct cttgcttaga gataccacct ttcccctgaa gtgttccttc
     3061 catgttttac ggcgagatgg tttctcctcg cctggccact cagccttagt tgtctctgtt
     3121 gtcttataga ggtctacttg aagaaggaaa aacagggggc atggtttgac tgtcctgtga
     3181 gcccttcttc cctgcctccc ccactcacag tgacccggaa tccctcgaca tggcagtcta
     3241 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3301 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3361 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgccag
     3421 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3481 tggcgaatgg aattccagac gattgagcgt caaaatgtag gtatttccat gagcgttttt
     3541 cctgttgcaa tggctggcgg taatattgtt ctggatatta ccagcaaggc cgatagtttg
     3601 agttcttcta ctcaggcaag tgatgttatt actaatcaaa gaagtattgc gacaacggtt
     3661 aatttgcgtg atggacagac tcttttactc ggtggcctca ctgattataa aaacacttct
     3721 caggattctg gcgtaccgtt cctgtctaaa atccctttaa tcggcctcct gtttagctcc
     3781 cgctctgatt ctaacgagga aagcacgtta tacgtgctcg tcaaagcaac catagtacgc
     3841 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3901 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3961 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4021 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4081 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4141 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4201 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4261 gaattttaac aaaatattaa cgtttacaat ttaaatattt gcttatacaa tcttcctgtt
     4321 tttggggctt ttctgattat caaccggggt acatatgatt gacatgctag ttttacgatt
     4381 accgttcatc gattctcttg tttgctccag actctcaggc aatgacctga tagcctttgt
     4441 agagacctct caaaaatagc taccctctcc ggcatgaatt tatcagctag aacggttgaa
     4501 tatcatattg atggtgattt gactgtctcc ggcctttctc acccgtttga atctttacct
     4561 acacattact caggcattgc atttaaaata tatgagggtt ctaaaaattt ttatccttgc
     4621 gttgaaataa aggcttctcc cgcaaaagta ttacagggtc ataatgtttt tggtacaacc
     4681 gatttagctt tatgctctga ggctttattg cttaattttg ctaattcttt gccttgcctg
     4741 tatgatttat tggatgttgg aattcctgat gcggtatttt ctccttacgc atctgtgcgg
     4801 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4861 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4921 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4981 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5041 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5161 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5221 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5281 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5341 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5401 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5461 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5521 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5581 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5641 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5701 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5761 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5821 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5881 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5941 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6001 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6061 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6121 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6181 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6241 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6301 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6361 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6421 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6481 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6541 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6601 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6661 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6721 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6781 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6841 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6901 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6961 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7021 gcctctcccc gcgcgttggc cgattcatta atgcagcagc tgcgcgctcg ctcgctcact
     7081 gaggccgccc
//
