LOCUS       sequence-305780-        6902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express LanCL1 in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6902)
  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 6902)
  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: pET-15b-LanCL1
FEATURES             Location/Qualifiers
     source          1..6902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     165..184
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        165..183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    184..208
                     /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)."
     RBS             223..245
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             264..281
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             291..308
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1567..1585)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1581..1628
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1803..1808,1809..1825,1826..1831))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1858..1876)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1944..2048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2049..2117,2118..2909)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2267..2286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3080..3668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3569..3588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3822..3839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3854..3993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3919..3938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4079..4101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4095..4286)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5598..6680)
                     /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     6642..6661
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6681..6758)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
       61 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
      121 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
      181 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa
      241 ggagatatac catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc
      301 gcggcagcca tatggctcaa agggccttcc cgaatcctta tgctgattat aacaaatccc
      361 tggccgaagg ctactttgat gctgccggga ggctgactcc tgagttctca caacgcttga
      421 ccaataagat tcgggagctt cttcagcaaa tggagagagg cctgaaatca gcagaccctc
      481 gggatggcac cggttacact ggctgggcag gtattgctgt gctttactta catctttatg
      541 atgtatttgg ggaccctgcc tacctacagt tagcacatgg ctatgtaaag caaagtctga
      601 actgcttaac caagcgctcc atcaccttcc tttgtgggga tgcaggcccc ctggcagtgg
      661 ccgctgtgct atatcacaag atgaacaatg agaagcaggc agaagattgc atcacacggc
      721 taattcacct aaataagatt gatcctcatg ctccaaatga aatgctctat gggcgaatag
      781 gctacatcta tgctcttctt tttgtcaata agaactttgg agtggaaaag attcctcaaa
      841 gccatattca gcagatttgt gaaacaattt taacctctgg agaaaaccta gctaggaaga
      901 gaaacttcac ggcaaagtct ccactgatgt atgaatggta ccaggaatat tatgtagggg
      961 ctgctcatgg cctggctgga atttattact acctgatgca gcccagcctt caagtgagcc
     1021 aagggaagtt acatagtttg gtcaagccca gtgtagacta cgtctgccag ctgaaattcc
     1081 cttctggcaa ttaccctcca tgtataggtg ataatcgaga tctgcttgtc cattggtgcc
     1141 atggcgcccc tggggtaatc tacatgctca tccaggccta taaggtattc agagaggaaa
     1201 agtatctctg tgatgcctat cagtgtgctg atgtgatctg gcaatatggg ttgctgaaga
     1261 agggatatgg gctgtgccac ggttctgcag ggaatgccta tgccttcctg acactctaca
     1321 acctcacaca ggacatgaag tacctgtata gggcctgtaa gtttgctgaa tggtgcttag
     1381 agtatggaga acatggatgc agaacaccag acaccccttt ctctctcttt gaaggaatgg
     1441 ctggaacaat atatttcctg gctgacctgc tagtccccac aaaagccagg ttccctgcat
     1501 ttgaactctg actcgaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1561 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1621 gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg
     1681 cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc
     1741 attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc
     1801 gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc
     1861 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1921 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1981 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2041 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2101 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2161 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2221 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2281 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2341 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2401 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2461 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2521 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2581 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2641 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2701 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2761 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2821 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2881 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2941 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3001 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3061 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3841 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3901 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3961 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4021 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4081 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     4141 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     4201 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     4261 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     4321 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     4381 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     4441 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     4501 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     4561 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4621 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4681 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     4741 atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag
     4801 atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc
     4861 aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt
     4921 ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa
     4981 ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat
     5041 aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg
     5101 atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg
     5161 cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc
     5221 gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg
     5281 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     5341 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     5401 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     5461 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     5521 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5581 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5641 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5701 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5761 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5821 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     5881 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5941 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     6001 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     6061 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6121 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     6181 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6241 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6301 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6361 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6421 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6481 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6541 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6601 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6661 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6721 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6781 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     6841 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     6901 ca
//
