LOCUS       sequence-196853-        7306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human transglutaminase 2 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7306)
  AUTHORS   Han BG, Cho JW, Cho YD, Jeong KC, Kim SY, Lee BI
  TITLE     Crystal structure of human transglutaminase 2 in complex with 
            adenosine triphosphate.
  JOURNAL   Int J Biol Macromol. 2010 Aug 1;47(2):190-5. doi: 
            10.1016/j.ijbiomac.2010.04.023. Epub 2010 May 5.
  PUBMED    20450932
REFERENCE   2  (bases 1 to 7306)
  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.ijbiomac.2010.04.023"; journalName: "Int J Biol 
            Macromol"; date: "2010-08-1- 1"; volume: "47"; issue: "2"; pages: 
            "190-5"
COMMENT     Sequence Label: pHis-hTG2
FEATURES             Location/Qualifiers
     source          1..7306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     180..199
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        180..198
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    199..223
                     /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             2330..2353
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2416..2434)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2430..2477
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2514..2969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2601..2620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2811..2832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3061..3876)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3061..3876)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3784..3803
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3868..3887
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3998..4586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4487..4506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4740..4757
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4772..4911
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4837..4856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4997..5019
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5013..5204)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6013..7095)
                     /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     7057..7076
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7096..7173)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
       61 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      121 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt
      181 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag atttaaatta
      241 actttaagaa ggagatatac atatggccga ggagctggtc ttagagaggt gtgatctgga
      301 gctggagacc aatggccgag accaccacac ggccgacctg tgccgggaga agctggtggt
      361 gcgacggggc cagcccttct ggctgaccct gcactttgag ggccgcaact acgaggccag
      421 tgtagacagt ctcaccttca gtgtcgtgac cggcccagcc cctagccagg aggccgggac
      481 caaggcccgt tttccactaa gagatgctgt ggaggagggt gactggacag ccaccgtggt
      541 ggaccagcaa gactgcaccc tctcgctgca gctcaccacc ccggccaacg cccccatcgg
      601 cctgtatcgc ctcagcctgg aggcctccac tggctaccag ggatccagct ttgtgctggg
      661 ccacttcatt ttgctcttca acgcctggtg cccagcggat gctgtgtacc tggactcgga
      721 agaggagcgg caggagtatg tcctcaccca gcagggcttt atctaccagg gctcggccaa
      781 gttcatcaag aacatacctt ggaattttgg gcagtttgaa gatgggatcc tagacatctg
      841 cctgatcctt ctagatgtca accccaagtt cctgaagaac gccggccgtg actgctcccg
      901 ccgcagcagc cccgtctacg tgggccgggt gggtagtggc atggtcaact gcaacgatga
      961 ccagggtgtg ctgctgggac gctgggacaa caactacggg gacggcgtca gccccatgtc
     1021 ctggatcggc agcgtggaca tcctgcggcg ctggaagaac cacggctgcc agcgcgtcaa
     1081 gtatggccag tgctgggtct tcgccgccgt ggcctgcaca gtgctgaggt gcctaggcat
     1141 ccctacccgc gtcgtgacca actacaactc ggcccatgac cagaacagca accttctcat
     1201 cgagtacttc cgcaatgagt ttggggagat ccagggtgac aagagcgaga tgatctggaa
     1261 cttccactgc tgggtggagt cgtggatgac caggccggac ctgcagccgg ggtacgaggg
     1321 ctggcaggcc ctggacccaa cgccccagga gaagagcgaa gggacgtact gctgtggccc
     1381 agttccagtt cgtgccatca aggagggcga cctgagcacc aagtacgatg cgccctttgt
     1441 ctttgcggag gtcaatgccg acgtggtaga ctggatccag caggacgatg ggtctgtgca
     1501 caaatccatc aaccgttccc tgatcgttgg gctgaagatc agcactaaga gcgtgggccg
     1561 agacgagcgg gaggatatca cccacaccta caaataccca gaggggtcct cagaggagag
     1621 ggaggccttc acaagggcga accacctgaa caaactggcc gagaaggagg agacagggat
     1681 ggccatgcgg atccgtgtgg gccagagcat gaacatgggc agtgactttg acgtctttgc
     1741 ccacatcacc aacaacaccg ctgaggagta cgtctgccgc ctcctgctct gtgcccgcac
     1801 cgtcagctac aatgggatct tggggcccga gtgtggcacc aagtacctgc tcaacctcaa
     1861 cctggagcct ttctctgaga agagcgttcc tctttgcatc ctctatgaga aataccgtga
     1921 ctgccttacg gagtccaacc tcatcaaggt gcgggccctc ctcgtggagc cagttatcaa
     1981 cagctacctg ctggctgagc gtgacctcta cctggagaat ccagaaatca agatccggat
     2041 ccttggggag cccaagcaga aacgcaagct ggtggctgag gtgtccctgc agaacccgct
     2101 ccctgtggcc ctggaaggct gcaccttcac tgtggagggg gccggcctga ctgaggagca
     2161 gaagacggtg gagatcccag accccgtgga ggcaggggag gaagttaagg tgagaatgga
     2221 cctgctgccg ctccacatgg gcctccacaa gctggtggtg aacttcgaga gcgacaagct
     2281 gaaggctgtg aagggcttcc ggaatgtcat cattggcccc gccctcgagc accaccacca
     2341 ccaccaccac cactaattga ttaataccta ggctgctaaa caaagcccga aaggaagctg
     2401 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     2461 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     2521 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2581 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2641 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2701 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2761 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2821 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2881 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2941 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3001 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     3061 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     3121 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     3181 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     3241 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     3301 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     3361 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     3421 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     3481 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     3541 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     3601 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     3661 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     3721 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     3781 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     3841 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     3901 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     3961 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4021 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4081 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4141 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4201 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4261 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4321 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4381 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4441 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4501 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4561 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4621 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4681 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4741 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4801 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4861 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4921 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4981 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5041 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     5101 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     5161 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     5221 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     5281 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     5341 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     5401 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     5461 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     5521 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     5581 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     5641 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     5701 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5761 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5821 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5881 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5941 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     6001 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6061 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     6121 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     6181 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     6241 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
     6301 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     6361 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     6421 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     6481 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     6541 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     6601 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     6661 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     6721 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     6781 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     6841 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6901 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6961 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     7021 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     7081 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     7141 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     7201 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     7261 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccac
//
