LOCUS       sequence-161649-        6844 bp    DNA     circular SYN 14-APR-2025
DEFINITION  MESA target chain with rapamycin FRB ectodomain, 20 extracellular 
            linkers, M cleavage sequence, and Gal4 transactivator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6844)
  AUTHORS   Hartfield RM, Schwarz KA, Muldoon JJ, Bagheri N, Leonard JN
  TITLE     Multiplexing Engineered Receptors for Multiparametric Evaluation of 
            Environmental Ligands.
  JOURNAL   ACS Synth Biol. 2017 Nov 17;6(11):2042-2055. doi: 
            10.1021/acssynbio.6b00279. Epub 2017 Aug 23.
  PUBMED    28771312
REFERENCE   2  (bases 1 to 6844)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.6b00279"; journalName: "ACS Synth Biol"; date: 
            "2017-11-17- 17"; volume: "6"; issue: "11"; pages: "2042-2055"
COMMENT     Sequence Label: Rap-MESA-20-M-Gal4
FEATURES             Location/Qualifiers
     source          1..6844
                     /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             1327..1347
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             1345..1785
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1936..2169
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     polyA_signal    2291..2683
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      3591..4104
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3678..3697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3888..3909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4585..4604)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4704..4726
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4764..4782)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4850..4954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4955..5023,5024..5815)
                     /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(5173..5192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5986..6574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6475..6494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6814..6844,1..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 agggatgctc gagatgtggc atgaaggcct ggaagaggca
      961 tctcgtttgt actttgggga aaggaacgtg aaaggcatgt ttgaggtgct ggagcccttg
     1021 catgctatga tggaacgggg cccccagact ctgaaggaaa catcctttaa tcaggcctat
     1081 ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt acatgaaatc agggaatgtc
     1141 aaggacctcc tccaagcctg ggacctctat tatcatgtgt tccgacgaat ctcaaagact
     1201 agaggcggat ctggaggctc tgggggttca ggcggatcag gaggcagtgg cgggtcaacc
     1261 ggtctggtcg tggttgctgg agtcctgttt tgttatggct tgctagtgac agtggctctt
     1321 tgtgttgaaa acctgtattt tcagatgaag ctactgtctt ctatcgaaca agcatgcgat
     1381 atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg
     1441 aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca
     1501 catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt
     1561 cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg
     1621 ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct
     1681 tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca
     1741 tcggaagaga gtagtaacaa aggtcaaaga cagttgactg taagcgctta cagccgcgcg
     1801 cgtacgaaaa acaattacgg gtctaccatc gagggcctgc tcgatctccc ggacgacgac
     1861 gcccccgaag aggcggggct ggcggctccg cgcctgtcct ttctccccgc gggacacacg
     1921 cgcagactgt cgacggcccc cccgaccgat gtcagcctgg gggacgagct ccacttagac
     1981 ggcgaggacg tggcgatggc gcatgccgac gcgctagacg atttcgatct ggacatgttg
     2041 ggggacgggg attccccggg tccgggattt accccccacg actccgcccc ctacggcgct
     2101 ctggatatgg ccgacttcga gtttgagcag atgtttaccg atgcccttgg aattgacgag
     2161 tacggtgggt aggcggccgc caccgcggtg gagctgttta aacctcgagc aattcacccc
     2221 accagtgcag gctgcctatc agaaagtggt ggctggtgtg gctaatgccc tggcccacaa
     2281 gtatcactaa gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa
     2341 gtccaactac taaactgggg atattatgaa gggccttgag catctggatt ctgcctaata
     2401 aaaaacattt attttcattg caatgatgta tttaaattat ttctgaatat tttactaaaa
     2461 agggaatgtg ggaggtcagt gcatttaaaa cataaagaaa tgaagagcta gttcaaacct
     2521 tgggaaaata cactatatct taaactccat gaaagaaggt gaggctgcaa acagtaatgc
     2581 acattggcaa cagcccctga tgcctatgcc ttattcatcc ctcagaaaag gattcaagta
     2641 gaggcttgat ttggaggtta aagttttgct atgctgtatt ttacattact tattgtttta
     2701 gctgtcctca tgaatgtctt ttcactaccc atttgcttat cctgcatctc tcagccttga
     2761 ctccactcag ttctcttgct tagagatacc acctttcccc tgaagtgttc cttccatgtt
     2821 ttacggcgag atggtttctc ctcgcctggc cactcagcct tagttgtctc tgttgtctta
     2881 tagaggtcta cttgaagaag gaaaaacagg gggcatggtt tgactgtcct gtgagccctt
     2941 cttccctgcc tcccccactc acagtgaccc ggaatccctc gacatggcag tctagagcat
     3001 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     3061 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3121 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg ccagctggcg
     3181 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3241 atggaattcc agacgattga gcgtcaaaat gtaggtattt ccatgagcgt ttttcctgtt
     3301 gcaatggctg gcggtaatat tgttctggat attaccagca aggccgatag tttgagttct
     3361 tctactcagg caagtgatgt tattactaat caaagaagta ttgcgacaac ggttaatttg
     3421 cgtgatggac agactctttt actcggtggc ctcactgatt ataaaaacac ttctcaggat
     3481 tctggcgtac cgttcctgtc taaaatccct ttaatcggcc tcctgtttag ctcccgctct
     3541 gattctaacg aggaaagcac gttatacgtg ctcgtcaaag caaccatagt acgcgccctg
     3601 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3661 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3721 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3781 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3841 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3901 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3961 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4021 taacaaaata ttaacgttta caatttaaat atttgcttat acaatcttcc tgtttttggg
     4081 gcttttctga ttatcaaccg gggtacatat gattgacatg ctagttttac gattaccgtt
     4141 catcgattct cttgtttgct ccagactctc aggcaatgac ctgatagcct ttgtagagac
     4201 ctctcaaaaa tagctaccct ctccggcatg aatttatcag ctagaacggt tgaatatcat
     4261 attgatggtg atttgactgt ctccggcctt tctcacccgt ttgaatcttt acctacacat
     4321 tactcaggca ttgcatttaa aatatatgag ggttctaaaa atttttatcc ttgcgttgaa
     4381 ataaaggctt ctcccgcaaa agtattacag ggtcataatg tttttggtac aaccgattta
     4441 gctttatgct ctgaggcttt attgcttaat tttgctaatt ctttgccttg cctgtatgat
     4501 ttattggatg ttggaattcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4561 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4621 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4681 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4741 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4801 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4921 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4981 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5041 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5101 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5161 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5221 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5281 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5341 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5401 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5461 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5521 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5581 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5641 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5701 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5761 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5821 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5881 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5941 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6001 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6061 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6121 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6601 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6661 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6721 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6781 ccccgcgcgt tggccgattc attaatgcag cagctgcgcg ctcgctcgct cactgaggcc
     6841 gccc
//
