LOCUS       sequence-166142-        7105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human tissue transglutaminase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  AUTHORS   Roy I, Smith O, Clouthier CM, Keillor JW
  TITLE     Expression, purification and kinetic characterisation of human 
            tissue transglutaminase.
  JOURNAL   Protein Expr Purif. 2013 Jan;87(1):41-6. doi: 
            10.1016/j.pep.2012.10.002. Epub 2012 Oct 22.
  PUBMED    23085087
REFERENCE   2  (bases 1 to 7105)
  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"; doi: 
            "10.1016/j.pep.2012.10.002"; journalName: "Protein Expr Purif"; 
            date: "2013-01"; volume: "87"; issue: "1"; pages: "41-6"
COMMENT     Sequence Label: pGST-PSP-hTG
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             921..944
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3155..3177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3322..3340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3408..3512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3513..3581,3582..4373)
                     /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(3731..3750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4544..5132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5033..5052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5286..5303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5376..5453
                     /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             5454..6536
                     /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(5473..5492)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6549..6570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6585..6590,6591..6608,6609..6615)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6623..6639
                     /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     6647..6663
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6647..6663
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6659..7105,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6679..6696)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6679..6695)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6688..6710)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg ctggaagttc tgttccaggg tccactggcc atggccgagg
      961 agctggtctt agagaggtgt gatctggagc tggagaccaa tggccgagac caccacacgg
     1021 ccgacctgtg ccgggagaag ctggtggtgc gacggggcca gcccttctgg ctgaccctgc
     1081 actttgaggg ccgcaactac gaggccagtg tagacagtct caccttcagt gtcgtgaccg
     1141 gcccagcccc tagccaggag gccgggacca aggcccgttt tccactaaga gatgctgtgg
     1201 aggagggtga ctggacagcc accgtggtgg accagcaaga ctgcaccctc tcgctgcagc
     1261 tcaccacccc ggccaacgcc cccatcggcc tgtatcgcct cagcctggag gcctccactg
     1321 gctaccaggg atccagcttt gtgctgggcc acttcatttt gctcttcaac gcctggtgcc
     1381 cagcggatgc tgtgtacctg gactcggaag aggagcggca ggagtatgtc ctcacccagc
     1441 agggctttat ctaccagggc tcggccaagt tcatcaagaa cataccttgg aattttgggc
     1501 agtttgaaga tgggatccta gacatctgcc tgatccttct agatgtcaac cccaagttcc
     1561 tgaagaacgc cggccgtgac tcgtcccggc gcagcagccc cgtctacgtg ggccgggtgg
     1621 gtagtggcat ggtcaactgc aacgatgacc agggtgtgct gctgggacgc tgggacaaca
     1681 actacgggga cggcgtcagc cccatgtcct ggatcggcag cgtggacatc ctgcggcgct
     1741 ggaagaacca cggctgccag cgcgtcaagt atggccagtg ctgggtcttc gccgccgtgg
     1801 cctgcacagt gctgaggtgc ctgggcatcc ctacccgcgt cgtgaccaac tacaactcgg
     1861 cccatgacca gaacagcaac cttctcatcg agtacttccg caatgagttt ggggagatcc
     1921 agggtgacaa gagcgagatg atctggaact tccactgctg ggtggagtcg tggatgacca
     1981 ggccggacct gcagccgggg tacgagggct ggcaggccct ggacccaacg ccccaggaga
     2041 agagcgaagg aacgtactgc tgtggcccag ttccagttcg tgccatcaag gagggcgacc
     2101 tgagcaccaa gtacgatgcg ccctttgtct ttgcggaggt caatgccgac gtggtagact
     2161 ggatccagca ggacgatggg tctgtgcaca aatccatcaa ccgttccctg atcgttgggc
     2221 tgaagatcag cactaagagc gtgggccgag acgagcggga ggatatcacc cacacctaca
     2281 aatacccaga ggggtcctca gaggagaggg aggccttcac aagggcgaac cacctgaaca
     2341 aactggccga gaaggaggag acagggatgg ccatgcggat ccgtgtgggc cagagcatga
     2401 acatgggcag tgactttgac gtctttgccc acatcaccaa caacaccgct gaggagtacg
     2461 tctgccgcct cctgctctgt gcccgcaccg tcagctacaa tgggatcttg gggcccgagt
     2521 gtggcaccaa gtacctgctc aacctcaacc tggagccttt ctctgagaag agcgttcctc
     2581 tttgcatcct ctatgagaaa taccgtgact gccttacgga gtccaacctc atcaaggtgc
     2641 gggccctcct cgtggagcca gttatcaaca gctacctgct ggctgagagg gacctctacc
     2701 tggagaatcc agaaatcaag atccggatcc ttggggagcc caagcagaaa cgcaagctgg
     2761 tggctgaggt gtccctgcag aacccgctcc ctgtggccct ggaaggctgc accttcactg
     2821 tggagggggc cggcctgact gaggagcaga agacggtgga gatcccagac cccgtggagg
     2881 caggggagga agttaaggtg agaatggacc tgctgccgct ccacatgggc ctccacaagc
     2941 tggtggtgaa cttcgagagc gacaagctga aggctgtgaa gggcttccgg aatgtcatca
     3001 ttggccccgc ctaagggacc cctgctccca gcctgctgag agcccccacc ttgatcccaa
     3061 tccttatccc aagcttcaga tctgctaagc ggtaccgaat tcatcgtgac tgactgacga
     3121 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3181 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3241 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     3301 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3361 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3421 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3481 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3541 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3601 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3661 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3721 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3781 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3841 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3901 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3961 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4021 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     4081 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4141 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4201 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4261 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4321 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4381 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4441 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4501 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4621 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4681 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4741 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4801 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4861 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4921 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4981 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5041 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5101 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5161 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5221 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5281 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     5341 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct
     5401 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     5461 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     5521 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     5581 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5641 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5701 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5761 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5821 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5881 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5941 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     6001 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     6061 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     6121 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     6181 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     6241 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     6301 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     6361 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     6421 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6481 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6541 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6601 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6661 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6721 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6781 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
     6841 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
     6901 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
     6961 caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
     7021 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
     7081 aattattttt gatggcgttg gaatt
//
