LOCUS       sequence-349920-        8604 bp    DNA     circular SYN 14-APR-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 Apr 14, 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        join(377..382,383..398,399..405)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    413..429
                     /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     418..440
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             508..525
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1474..1483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1591..1614
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1627..1651
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1672..1692
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3207..3218
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3632..3656)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3668..3685)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3668..3684)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3677..3699)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3863..3880)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4033..4119
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4211..4238
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4257..4348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4349..4417,4418..5209)
                     /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(4567..4586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5251..5764)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5446..5467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5658..5677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      5875..6463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6364..6383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6617..6634
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6649..6789
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6715..6734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6875..6897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6891..7082)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        7578..7655
                     /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             join(7656..8604,1..134)
                     /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(7675..7694)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
       61 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      121 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc
      181 tcatgtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
      241 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
      301 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
      361 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
      421 ggataacaat ttcacacagg aaacagccag tccgtttagg tgttttcacg agcacttcac
      481 caacaaggac catagattat gaaaatccat caccatcacc atcacgaaga aggtaaactg
      541 gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc
      601 gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc
      661 ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt
      721 ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac
      781 aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg
      841 atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa
      901 acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg
      961 atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat
     1021 gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc
     1081 gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac
     1141 accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac
     1201 ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg
     1261 ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac
     1321 gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat
     1381 gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac
     1441 gaggaagagt tggcgaaaga tccacgtatt gccgccacca tggaaaacgc ccagaaaggt
     1501 gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg
     1561 atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactaat
     1621 tcgatcacaa gtttgtacaa aaaagcaggc tccgcggccg cccccttcac cgagaacttg
     1681 tacttccaag gtatggtgag caaaggtgaa gaactgttta ccggtgttgt tccgattctg
     1741 gttgaactgg atggtgacgt taatggtcac aaattttcag ttagcggtga aggcgaaggt
     1801 gatgcaacct atggtaaact gaccctgaaa tttatctgta ccaccggcaa actgccggtt
     1861 ccgtggccga ccctggttac caccctgagc tggggtgttc agtgttttgc acgttatccg
     1921 gatcacatga aacagcacga ttttttcaaa agcgcaatgc cggaaggtta tgttcaagaa
     1981 cgtaccatct tcttcaaaga tgacggcaac tataaaaccc gtgccgaagt taaatttgaa
     2041 ggtgataccc tggtgaatcg cattgaactg aaaggcatcg attttaaaga ggatggtaat
     2101 atcctgggcc acaaactgga atataatgcc attcatggca acgtgtatat caccgcagat
     2161 aaacagaaaa acggcatcaa agcaaatttt ggcctgaact gcaatattga agatggttca
     2221 gttcagctgg cagatcatta tcagcagaat accccgattg gtgatggtcc ggttctgctg
     2281 ccggataatc attatctgag cacccagagc aaactgagca aagatccgaa tgaaaaacgt
     2341 gatcacatgg tgctgctgga atttgttacc gcagcaggta ttaccctggg tatggatgaa
     2401 ctgtataaag agctcatgca gtcgatgaaa gaaacagctt cgaatattgc agcttctgca
     2461 aaatctggca tggacaaaac caaagctacc ttggaggaaa aggcggagaa gatgaagaca
     2521 cgagaccctg ttcagaaaca gatggctaca caggttaaag aagataagat caatcaagct
     2581 gagatgcaga agagagaaac gcgtcagcac aacgcggcca tgaaagaagc ggctggagcc
     2641 ggaaccggtt taggtttggg gacggccact cactcgacca ctggacaagt cggacacggc
     2701 actgggacac atcagatgtc ggctctgcct ggtcacggaa cgggacaact gaccgaccgc
     2761 gttgtggagg gcacggctgt gaccgacccg attggaagga acactggaac tggtcggaca
     2821 accgctcata acacacacgt tggtggtggt ggtgccaccg ggtacggaac cggcggggga
     2881 tatactggaa gatctatggt gtcaaaaggt gaggaactgt ttacaggcgt ggtgccgatc
     2941 ctggtagagc tggacgggga tgtgaatggc cataaattca gcgtttcagg tgaaggtgag
     3001 ggcgacgcca cgtacggaaa actgacactg aaattcattt gcacaacagg taaactgcct
     3061 gtgccttggc ctacactggt gaccaccttt ggttatggtc tgatgtgctt tgctcgctat
     3121 cctgaccaca tgaaacaaca tgatttcttt aaatctgcca tgcctgaagg ctacgtgcaa
     3181 gagcgcacca tttttttcaa agacgatggg aattacaaaa cacgtgcgga ggtgaaattc
     3241 gagggcgata cactggttaa ccgtatcgag ctgaaaggta tcgacttcaa agaggacgga
     3301 aacattctgg gtcataaact ggaatacaac tacaacagcc ataacgtgta catcatggcc
     3361 gacaaacaaa aaaacgggat taaagtgaac ttcaaaatcc gccacaacat cgaagatggc
     3421 agcgtgcagc tggccgacca ctatcaacaa aacacaccga tcggcgacgg tcctgtactg
     3481 ctgcctgaca accactatct gtcatatcag agcgcactgt caaaagatcc taacgagaaa
     3541 cgcgaccaca tggttctgct ggaattcgtg acagccgctg gcattacact gggcatggac
     3601 gagctgtaca aataaaaggg tgggcgcgcc gacccagctt tcttgtacaa agtggtgata
     3661 gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     3721 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     3781 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcagctt ggctgttttg
     3841 gcggatgaga taagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     3901 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     3961 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     4021 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     4081 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     4141 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     4201 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     4261 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4321 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4381 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4441 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4501 cggtaagatc cttgagagtt ttcgccccga agaacgtttc ccaatgatga gcacttttaa
     4561 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4621 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4681 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4741 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4801 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4861 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4921 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4981 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5041 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5101 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5161 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5221 agtttactca tatatacttt agattgattt accccggttg ataatcagaa aagccccaaa
     5281 aacaggaaga ttgtataagc aaatatttaa attgtaaacg ttaatatttt gttaaaattc
     5341 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5401 ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag
     5461 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5521 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     5581 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5641 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5701 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5761 gcgtaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5821 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6001 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6541 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6601 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     6661 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     6721 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     6781 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6841 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6901 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc
     6961 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     7021 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     7081 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     7141 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     7201 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     7261 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     7321 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     7381 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     7441 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     7501 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca
     7561 acgctgcccg aaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
     7621 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     7681 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     7741 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     7801 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     7861 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     7921 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     7981 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     8041 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     8101 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     8161 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     8221 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     8281 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     8341 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     8401 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     8461 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     8521 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     8581 agggccaggc ggtgaagggc aatc
//
