LOCUS       sequence-305773-        7986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express MBP-LanCL2 in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7986)
  AUTHORS   Lai KY, Galan SRG, Zeng Y, Zhou TH, He C, Raj R, Riedl J, Liu S, 
            Chooi KP, Garg N, Zeng M, Jones LH, Hutchings GJ, Mohammed S, Nair 
            SK, Chen J, Davis BG, van der Donk WA
  TITLE     LanCLs add glutathione to dehydroamino acids generated at 
            phosphorylated sites in the proteome.
  JOURNAL   Cell. 2021 Apr 27. pii: S0092-8674(21)00436-0. doi: 
            10.1016/j.cell.2021.04.001.
  PUBMED    33932340
REFERENCE   2  (bases 1 to 7986)
  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: "Cell. 2021 
            Apr 27. pii: S0092-8674(21)00436-0. doi: 
            10.1016/j.cell.2021.04.001."
COMMENT     Sequence Label: pMAL-p2x-LanCL2
FEATURES             Location/Qualifiers
     source          1..7986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(725..730,731..746,747..753)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    761..777
                     /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     766..788
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             847..1947
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1921..1944
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1996..2007
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3396..3413)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3396..3412)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3405..3427)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3591..3608)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3761..3847
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3939..3966
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3985..4076
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4077..4145,4146..4937)
                     /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(4295..4314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4979..5492)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5174..5195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5386..5405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      5603..6191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6092..6111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6345..6362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6377..6519
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6445..6464)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6605..6627
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6621..6812)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        7308..7385
                     /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(7386..7986,1..482)
                     /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(7405..7424)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
       61 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      121 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      181 ggctgcgcgt tggtgcggat atctcagtag tgggatacga cgataccgaa gacagctcat
      241 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      301 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
      361 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gagcgcaacg caattaatgt aagttagctc actcattagg cacaattctc atgtttgaca
      541 gcttatcatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg
      601 tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc
      661 gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga
      721 gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt
      781 cacacaggaa acagccagtc cgtttaggtg ttttcacgag cacttcacca acaaggacca
      841 tagcatatga aaatcgaaga aggtaaactg gtaatctgga ttaacggcga taaaggctat
      901 aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa agtcaccgtt
      961 gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg cgatggccct
     1021 gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg cctgttggct
     1081 gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg ggatgccgta
     1141 cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc gctgatttat
     1201 aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc gctggataaa
     1261 gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc
     1321 tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg caagtacgac
     1381 attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt cctggttgac
     1441 ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga agctgccttt
     1501 aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa catcgacacc
     1561 agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc atccaaaccg
     1621 ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga gctggcaaaa
     1681 gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa taaagacaaa
     1741 ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga tccacgtatt
     1801 gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc
     1861 gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg tcagactgtc
     1921 gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa caataacaat
     1981 aacaacaacc tcgggatcga gggaaggatt tcagaattca tgggcgagac catgtcaaag
     2041 aggctgaagc tccacctggg aggggaggca gaaatggagg aacgggcgtt cgtcaacccc
     2101 ttcccggact acgaggccgc cgccggggcg ctgctcgcct ccggagcggc cgaagagaca
     2161 ggctgtgttc gtcccccggc caccccggat gagcccggcc tcccttttca tcaggacggg
     2221 aagatcattc ataatttcat aagacggatc cagaccaaaa ttaaagatct tctgcagcaa
     2281 atggaagaag ggctgaagac agctgatccc catgactgct ctgcttatac tggctggaca
     2341 ggcatagccc ttttgtacct gcagttgtac cgggtcacat gtgaccaaac ctacctgctc
     2401 cgatccctgg attacgtaaa aagaacactt cggaatctga atggccgcag ggtcaccttc
     2461 ctctgtgggg atgctggccc cctggctgtt ggagctgtga tttatcacaa actcagaagt
     2521 gactgtgagt cccaggaatg tgtcacaaaa cttttgcagc tccagagatc ggttgtctgc
     2581 caagaatcag accttcctga tgagctgctt tatggacggg caggttatct gtatgcctta
     2641 ctgtacctga acacagagat aggtccaggc accgtgtgtg agtcagctat taaagaggta
     2701 gtcaatgcta ttattgaatc gggtaagact ttgtcaaggg aagaaagaaa aacggagcgc
     2761 tgcccgctgt tgtaccagtg gcaccggaag cagtacgttg gagcagccca tggcatggct
     2821 ggaatttact atatgttaat gcagccggca gcaaaagtgg accaagaaac cttgacagaa
     2881 atggtgaaac ccagtattga ttatgtgcgc cacaaaaaat tccgatctgg gaattaccca
     2941 tcatcattaa gcaatgaaac agaccggctg gtgcactggt gccacggcgc cccgggggtc
     3001 atccacatgc tcatgcaggc gtacaaggtc tttaaggagg agaagtactt gaaagaggcc
     3061 atggagtgta gcgatgtgat ttggcagcga ggtttgctgc ggaagggcta cgggatatgc
     3121 catgggactg ctggcaacgg ctattccttc ctgtcccttt accgtctcac gcaggataag
     3181 aagtacctct accgagcttg caagtttgca gagtggtgtc tagattacgg agcacacggg
     3241 tgccgcattc ctgacagacc ctattcgctc tttgaaggca tggctggcgc tattcacttt
     3301 ctctctgatg tcctgggacc agagacatca cggtttccag catttgaact tgactcttcg
     3361 aagagggatt aactcgacct gcaggcaagc ttggcactgg ccgtcgtttt acaacgtcgt
     3421 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3481 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3541 aatggcgaat ggcagcttgg ctgttttggc ggatgagata agattttcag cctgatacag
     3601 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     3661 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     3721 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     3781 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     3841 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     3901 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     3961 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
     4021 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4081 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4141 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4201 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4261 aacgtttccc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     4321 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4381 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4441 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4501 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4561 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4621 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4681 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4741 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4801 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4861 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4921 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac
     4981 cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat
     5041 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5101 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5161 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5221 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     5281 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5341 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5401 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5461 cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat ctaggtgaag atcctttttg
     5521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6301 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6361 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6421 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     6481 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     6541 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6601 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     6661 ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc ctgttcatcc
     6721 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     6781 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     6841 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     6901 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     6961 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     7021 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     7081 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     7141 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     7201 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     7261 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg aaattccgac accatcgaat
     7321 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     7381 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     7441 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     7501 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     7561 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     7621 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     7681 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     7741 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     7801 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     7861 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     7921 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     7981 ctggct
//
