LOCUS       sequence-311995-        5528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of rhomboid protease EcGlpG without N terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  AUTHORS   Mihaljevic L, Urban S
  TITLE     Decoding the Functional Evolution of an Intramembrane Protease 
            Superfamily by Statistical Coupling Analysis.
  JOURNAL   Structure. 2020 Aug 10. pii: S0969-2126(20)30279-3. doi: 
            10.1016/j.str.2020.07.015.
  PUBMED    32795403
REFERENCE   2  (bases 1 to 5528)
  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: "Structure. 
            2020 Aug 10. pii: S0969-2126(20)30279-3. doi: 
            10.1016/j.str.2020.07.015."
COMMENT     Sequence Label: pGEX-delN-EcGlpG
FEATURES             Location/Qualifiers
     source          1..5528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..759
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1396..1418)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1563..1581)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1649..1753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1754..1822,1823..2614)
                     /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(1972..1991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2785..3373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3274..3293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3527..3544
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3617..3694
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3695..4777
                     /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(3714..3733)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4826..4831,4832..4849,4850..4856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4864..4880
                     /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     4888..4904
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4888..4904
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4900..5427
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4920..4937)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4920..4936)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4929..4951)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctgga agttctgttc caggggccca tggccgcctt gcgtgaacgc
      781 gcaggtccgg taacctgggt gatgatgatc gcctgcgtgg tggtgtttat tgccatgcaa
      841 attctcggcg atcaggaagt gatgttatgg ctggcctggc cattcgatcc aacactgaaa
      901 tttgagttct ggcgttactt cacccacgcg ttaatgcact tctcgctgat gcatatcctc
      961 tttaacctgc tctggtggtg gtatctcggc ggtgcggtgg aaaaacgcct cggtagcggt
     1021 aagctaattg tcattacgct tatcagcgcc ctgttaagcg gctatgtgca gcaaaaattc
     1081 agcgggccgt ggtttggcgg gctttctggc gtggtgtatg cgctgatggg ctacgtctgg
     1141 ctacgtggcg aacgcgatcc gcaaagtggc atttacctgc aacgtgggtt aattatcttt
     1201 gcgctgatct ggattgtcgc cggatggttt gatttgtttg ggatgtcgat ggcgaacgga
     1261 gcacacatcg ccgggttagc cgtgggttta gcgatggctt ttgttgattc gctcaatgcg
     1321 cgaaaacgaa aataagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg
     1381 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1441 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1501 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa
     1561 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1621 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1681 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1741 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1801 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1861 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1921 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1981 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2041 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2101 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2161 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2221 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2281 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2341 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2401 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2461 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2521 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2581 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2641 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2701 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3421 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3481 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3541 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3601 acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     3661 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     3721 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     3781 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3841 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3901 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3961 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     4021 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     4081 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     4141 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     4201 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4261 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4321 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4381 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     4441 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     4501 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     4561 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     4621 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     4681 gctagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4741 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     4801 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4861 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca
     4921 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4981 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     5041 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
     5101 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
     5161 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc
     5221 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
     5281 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
     5341 ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
     5401 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
     5461 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
     5521 atgagctg
//
