LOCUS       sequence-345843-        8604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SED1 biosensor for expression in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8604)
  AUTHORS   Cuevas-Velazquez CL, Vellosillo T, Guadalupe K, Schmidt HB, Yu F, 
            Moses D, Brophy JAN, Cosio-Acosta D, Das A, Wang L, Jones AM, 
            Covarrubias AA, Sukenik S, Dinneny JR
  TITLE     Intrinsically disordered protein biosensor tracks the 
            physical-chemical effects of osmotic stress on cells.
  JOURNAL   Nat Commun. 2021 Sep 14;12(1):5438. doi: 10.1038/s41467-021-25736-8.
  PUBMED    34521831
REFERENCE   2  (bases 1 to 8604)
  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 
            Commun."; date: "2021-09-14"; volume: "12(1):5438. doi"; pages: " 
            10.1038/s41467-021-25736-8"
COMMENT     Sequence Label: pDEST-HisMBP-SED1
FEATURES             Location/Qualifiers
     source          1..8604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..80
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             81..1163
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(100..119)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1406..1411,1412..1427,1428..1434)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1447..1469
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1537..1554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2503..2512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2620..2643
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    2656..2680
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2701..2721
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4236..4247
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(4661..4685)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4697..4714)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4697..4713)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4706..4728)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4892..4909)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5062..5148
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5240..5267
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5286..5377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5378..5446,5447..6238)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5596..5615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6280..6793)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6475..6496)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6687..6706
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      6904..7492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7393..7412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7646..7663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7678..7818
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7744..7763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7904..7926
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7920..8111)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcacttcacc aacaaggacc atagattatg aaaatccatc accatcacca tcacgaagaa
     1561 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
     1621 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
     1681 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
     1741 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
     1801 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
     1861 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
     1921 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
     1981 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
     2041 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     2101 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     2161 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     2221 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     2281 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     2341 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     2401 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     2461 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccaccat ggaaaacgcc
     2521 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     2581 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     2641 cagactaatt cgatcacaag tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc
     2701 gagaacttgt acttccaagg tatggtgagc aaaggtgaag aactgtttac cggtgttgtt
     2761 ccgattctgg ttgaactgga tggtgacgtt aatggtcaca aattttcagt tagcggtgaa
     2821 ggcgaaggtg atgcaaccta tggtaaactg accctgaaat ttatctgtac caccggcaaa
     2881 ctgccggttc cgtggccgac cctggttacc accctgagct ggggtgttca gtgttttgca
     2941 cgttatccgg atcacatgaa acagcacgat tttttcaaaa gcgcaatgcc ggaaggttat
     3001 gttcaagaac gtaccatctt cttcaaagat gacggcaact ataaaacccg tgccgaagtt
     3061 aaatttgaag gtgataccct ggtgaatcgc attgaactga aaggcatcga ttttaaagag
     3121 gatggtaata tcctgggcca caaactggaa tataatgcca ttcatggcaa cgtgtatatc
     3181 accgcagata aacagaaaaa cggcatcaaa gcaaattttg gcctgaactg caatattgaa
     3241 gatggttcag ttcagctggc agatcattat cagcagaata ccccgattgg tgatggtccg
     3301 gttctgctgc cggataatca ttatctgagc acccagagca aactgagcaa agatccgaat
     3361 gaaaaacgtg atcacatggt gctgctggaa tttgttaccg cagcaggtat taccctgggt
     3421 atggatgaac tgtataaaga gctcatgcag tcgatgaaag aaacagcttc gaatattgca
     3481 gcttctgcaa aatctggcat ggacaaaacc aaagctacct tggaggaaaa ggcggagaag
     3541 atgaagacac gagaccctgt tcagaaacag atggctacac aggttaaaga agataagatc
     3601 aatcaagctg agatgcagaa gagagaaacg cgtcagcaca acgcggccat gaaagaagcg
     3661 gctggagccg gaaccggttt aggtttgggg acggccactc actcgaccac tggacaagtc
     3721 ggacacggca ctgggacaca tcagatgtcg gctctgcctg gtcacggaac gggacaactg
     3781 accgaccgcg ttgtggaggg cacggctgtg accgacccga ttggaaggaa cactggaact
     3841 ggtcggacaa ccgctcataa cacacacgtt ggtggtggtg gtgccaccgg gtacggaacc
     3901 ggcgggggat atactggaag atctatggtg tcaaaaggtg aggaactgtt tacaggcgtg
     3961 gtgccgatcc tggtagagct ggacggggat gtgaatggcc ataaattcag cgtttcaggt
     4021 gaaggtgagg gcgacgccac gtacggaaaa ctgacactga aattcatttg cacaacaggt
     4081 aaactgcctg tgccttggcc tacactggtg accacctttg gttatggtct gatgtgcttt
     4141 gctcgctatc ctgaccacat gaaacaacat gatttcttta aatctgccat gcctgaaggc
     4201 tacgtgcaag agcgcaccat ttttttcaaa gacgatggga attacaaaac acgtgcggag
     4261 gtgaaattcg agggcgatac actggttaac cgtatcgagc tgaaaggtat cgacttcaaa
     4321 gaggacggaa acattctggg tcataaactg gaatacaact acaacagcca taacgtgtac
     4381 atcatggccg acaaacaaaa aaacgggatt aaagtgaact tcaaaatccg ccacaacatc
     4441 gaagatggca gcgtgcagct ggccgaccac tatcaacaaa acacaccgat cggcgacggt
     4501 cctgtactgc tgcctgacaa ccactatctg tcatatcaga gcgcactgtc aaaagatcct
     4561 aacgagaaac gcgaccacat ggttctgctg gaattcgtga cagccgctgg cattacactg
     4621 ggcatggacg agctgtacaa ataaaagggt gggcgcgccg acccagcttt cttgtacaaa
     4681 gtggtgatag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4741 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4801 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcagcttg
     4861 gctgttttgg cggatgagat aagattttca gcctgataca gattaaatca gaacgcagaa
     4921 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     4981 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     5041 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     5101 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     5161 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     5221 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     5281 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5341 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5401 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5461 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5521 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag
     5581 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     5641 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5701 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5761 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5821 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5881 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5941 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6001 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6061 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6121 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6181 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6241 gtcagaccaa gtttactcat atatacttta gattgattta ccccggttga taatcagaaa
     6301 agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt taatattttg
     6361 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6421 ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt
     6481 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6541 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
     6601 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6661 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6721 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6781 ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6841 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6901 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6961 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7021 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     7081 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7141 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7201 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7261 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7321 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7381 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7441 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7501 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7561 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7621 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     7681 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     7741 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     7801 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     7861 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     7921 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     7981 tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     8041 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     8101 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     8161 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     8221 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     8281 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     8341 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     8401 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     8461 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     8521 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     8581 caggacccaa cgctgcccga aatt
//
