LOCUS       sequence-496821-        5420 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  N-term GST tag E2 variant selected for LUBAC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5420)
  TITLE     Schulman lab plasmids 2025
REFERENCE   2  (bases 1 to 5420)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: GST-tag LUBAC binding E2 variant L3LU-1
FEATURES             Location/Qualifiers
     source          1..5420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..654
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     612..634
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             661..681
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1213..1235)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1380..1398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1466..1570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1571..1639,1640..2431)
                     /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(1789..1808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2602..3190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3091..3110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3344..3361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3434..3511
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3512..4594
                     /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(3531..3550)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4643..4648,4649..4666,4667..4673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4681..4697
                     /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     4705..4721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4705..4721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4717..5244
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4737..4754)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4737..4753)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4746..4768)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5346..5351,5352..5367,5368..5374)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5382..5398
                     /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)."
ORIGIN
        1 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
       61 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
      121 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
      181 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
      241 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
      301 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
      361 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
      421 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
      481 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
      541 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
      601 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat
      661 gaaaacctgt attttcaggg atccatgcgt gcggcctgga ggaggctgcc gcgtgagctt
      721 gaagaaatcc gcaaagcagg gatgaaaaac ttccgtaaca tccaggttga tgaagctaat
      781 ttattgactt ggcaagggct tattgttcct gacaaccctc catatgataa gggagccttc
      841 agaatcgaaa tcaactttcc agcagagtac ccattcaaac caccgaagat cacatttaaa
      901 acaaagatct atcacccaaa catcgacgaa aaggggcagg tctgtctgcc agtaattagt
      961 gccgaaaact ggaagccagc aaccaaaacc gaccaagtaa tccagtccct catagcactg
     1021 gtgaatgacc cccagcctga gcacccgctt cgggctgacc tagctgaaga atactctaag
     1081 gaccgtaaaa aattcgcaaa gaatgctgaa gagtttacaa agaaatatgg ggaaaagcga
     1141 cctgtggact aagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
     1201 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1261 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1321 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
     1381 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1801 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2101 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2881 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcacaca
     2941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3361 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3421 ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     3481 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     3541 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     3601 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     3661 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     3721 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     3781 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     3841 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     3901 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     3961 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     4021 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     4081 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     4141 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     4201 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     4261 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     4321 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     4381 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     4441 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     4501 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4561 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     4621 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     4681 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
     4741 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4801 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4861 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     4921 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     4981 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     5041 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     5101 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga
     5161 attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct
     5221 gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc
     5281 cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg
     5341 agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt
     5401 tcacacagga aacagtattc
//
