LOCUS       sequence-94231-e        7474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA protease chain with CD4 ectodomain and no linker domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7474)
  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 7474)
  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: CD4_LD 0_TEV
FEATURES             Location/Qualifiers
     source          1..7474
                     /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             2095..2799
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     polyA_signal    2921..3313
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      4221..4734
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4308..4327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4518..4539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5215..5234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5334..5356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5394..5412)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5480..5584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5585..5653,5654..6445)
                     /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(5803..5822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6616..7204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7105..7124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7444..7474,1..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 agggaagacg ctggtgctgg ggaaggaagg ggaatcagca
      961 gaactgccct gcgagagttc ccagaagaag atcacagtct tcacctggaa gttctctgac
     1021 cagaggaaga ttctggggca gcatggcaaa ggtgtattaa ttagaggagg ttcgccttcg
     1081 cagtttgatc gttttgattc caaaaaaggg gcatgggaga aaggatcgtt tcctctcatc
     1141 atcaataaac ttaagatgga agactctcag acttatatct gtgagctgga gaacaggaaa
     1201 gaggaggtgg agttgtgggt gttcaaagtg accttcagtc cgggtaccag cctgttgcaa
     1261 gggcagagcc tgaccctgac cttggatagc aactctaagg tctctaaccc cttgacagag
     1321 tgcaaacaca aaaagggtaa agttgtcagt ggttccaaag ttctctccat gtccaaccta
     1381 agggttcagg acagcgactt ctggaactgc accgtgaccc tggaccagaa aaagaactgg
     1441 ttcggcatga cactctcagt gctgggtttt cagagcacag ctatcacggc ctataagagt
     1501 gagggagagt cagcggagtt ctccttccca ctcaactttg cagaggaaaa cgggtgggga
     1561 gagctgatgt ggaaggcaga gaaggattct ttcttccagc cctggatctc cttctccata
     1621 aagaacaaag aggtgtccgt acaaaagtcc accaaagacc tcaagctcca gctgaaggaa
     1681 acgctcccac tcaccctcaa gataccccag gtctcgcttc agtttgctgg ttctggcaac
     1741 ctgactctga ctctggacaa agggacactg catcaggaag tgaacctggt ggtgatgaaa
     1801 gtggctcagc tcaacaatac tttgacctgt gaggtgatgg gacctacctc tcccaagatg
     1861 agactgaccc tgaagcagga gaaccaggag gccagggtct ctgaggagca gaaagtagtt
     1921 caagtggtgg cccctgagac agggctgtgg cagtgtctac tgagtgaagg tgataaggtc
     1981 aagatggact ccaggatcca ggttttatcc agagggaagc ttttttgggc actggtcgtg
     2041 gttgctggag tcctgttttg ttatggcttg ctagtgacag tggctctttg tgttgagagc
     2101 ttgtttaagg ggccgcgtga ttacaacccg atatcgagca ccatttgtca tttgacgaat
     2161 gaatctgatg ggcacacaac atcgttgtat ggtattggat ttggtccctt catcattaca
     2221 aacaagcact tgtttagaag aaataatgga acactgttgg tccaatcact acatggtgta
     2281 ttcaaggtca agaacaccac gactttgcaa caacacctca ttgatgggag ggacatgata
     2341 attattcgca tgcctaagga tttcccacca tttcctcaaa agctgaaatt tagagagcca
     2401 caaagggaag agcgcatatg tcttgtgaca accaacttcc aaactaagag catgtctagc
     2461 atggtgtcag acactagttg cacattccct tcatctgatg gcatattctg gaagcattgg
     2521 attcaaacca aggatgggca gtgtggcagt ccattagtat caactagaga tgggttcatt
     2581 gttggtatac actcagcatc gaatttcacc aacacaaaca attatttcac aagcgtgccg
     2641 aaaaacttca tggaattgtt gacaaatcag gaggcgcagc agtgggttag tggttggcga
     2701 ttaaatgctg actcagtatt gtgggggggc cataaagttt tcatggtgaa acctgaagag
     2761 ccttttcagc cagttaagga agcgactcaa ctcatgaatt aggcggccgc caccgcggtg
     2821 gagctgttta aacctcgagc aattcacccc accagtgcag gctgcctatc agaaagtggt
     2881 ggctggtgtg gctaatgccc tggcccacaa gtatcactaa gctcgctttc ttgctgtcca
     2941 atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg atattatgaa
     3001 gggccttgag catctggatt ctgcctaata aaaaacattt attttcattg caatgatgta
     3061 tttaaattat ttctgaatat tttactaaaa agggaatgtg ggaggtcagt gcatttaaaa
     3121 cataaagaaa tgaagagcta gttcaaacct tgggaaaata cactatatct taaactccat
     3181 gaaagaaggt gaggctgcaa acagtaatgc acattggcaa cagcccctga tgcctatgcc
     3241 ttattcatcc ctcagaaaag gattcaagta gaggcttgat ttggaggtta aagttttgct
     3301 atgctgtatt ttacattact tattgtttta gctgtcctca tgaatgtctt ttcactaccc
     3361 atttgcttat cctgcatctc tcagccttga ctccactcag ttctcttgct tagagatacc
     3421 acctttcccc tgaagtgttc cttccatgtt ttacggcgag atggtttctc ctcgcctggc
     3481 cactcagcct tagttgtctc tgttgtctta tagaggtcta cttgaagaag gaaaaacagg
     3541 gggcatggtt tgactgtcct gtgagccctt cttccctgcc tcccccactc acagtgaccc
     3601 ggaatccctc gacatggcag tctagagcat ggctacgtag ataagtagca tggcgggtta
     3661 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3721 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3781 tcagtgagcg agcgagcgcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3841 ttcccaacag ttgcgcagcc tgaatggcga atggaattcc agacgattga gcgtcaaaat
     3901 gtaggtattt ccatgagcgt ttttcctgtt gcaatggctg gcggtaatat tgttctggat
     3961 attaccagca aggccgatag tttgagttct tctactcagg caagtgatgt tattactaat
     4021 caaagaagta ttgcgacaac ggttaatttg cgtgatggac agactctttt actcggtggc
     4081 ctcactgatt ataaaaacac ttctcaggat tctggcgtac cgttcctgtc taaaatccct
     4141 ttaatcggcc tcctgtttag ctcccgctct gattctaacg aggaaagcac gttatacgtg
     4201 ctcgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4261 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4321 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4381 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4441 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4501 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4561 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4621 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaat
     4681 atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg gggtacatat
     4741 gattgacatg ctagttttac gattaccgtt catcgattct cttgtttgct ccagactctc
     4801 aggcaatgac ctgatagcct ttgtagagac ctctcaaaaa tagctaccct ctccggcatg
     4861 aatttatcag ctagaacggt tgaatatcat attgatggtg atttgactgt ctccggcctt
     4921 tctcacccgt ttgaatcttt acctacacat tactcaggca ttgcatttaa aatatatgag
     4981 ggttctaaaa atttttatcc ttgcgttgaa ataaaggctt ctcccgcaaa agtattacag
     5041 ggtcataatg tttttggtac aaccgattta gctttatgct ctgaggcttt attgcttaat
     5101 tttgctaatt ctttgccttg cctgtatgat ttattggatg ttggaattcc tgatgcggta
     5161 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     5221 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5281 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5341 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5401 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5581 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5641 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5701 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5761 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5821 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5881 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5941 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6001 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6061 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6121 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6181 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6241 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6301 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6361 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6421 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6481 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6541 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6601 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6661 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6721 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6781 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6841 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6901 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6961 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7021 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7081 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7141 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7201 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7261 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7321 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7381 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     7441 cagctgcgcg ctcgctcgct cactgaggcc gccc
//
