LOCUS       sequence-220214-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the PEST domain-replaced 48 kDa form of human dematin with a
            cleavable N-terminal GST tag and non-cleavable C-terminal 6-his tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  AUTHORS   Chen L, Brown JW, Mok YF, Hatters DM, McKnight CJ
  TITLE     The allosteric mechanism induced by protein kinase A (PKA) 
            phosphorylation of dematin (band 4.9).
  JOURNAL   J Biol Chem. 2013 Mar 22;288(12):8313-20. doi: 
            10.1074/jbc.M112.438861. Epub 2013 Jan 25.
  PUBMED    23355471
REFERENCE   2  (bases 1 to 6145)
  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: 
            "10.1074/jbc.M112.438861"; journalName: "J Biol Chem"; date: 
            "2013-03-22- 22"; volume: "288"; issue: "12"; pages: "8313-20"
COMMENT     Sequence Label: rd-his
FEATURES             Location/Qualifiers
     source          1..6145
                     /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..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             754..777
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1927..1944
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2013..2035)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2180..2198)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2266..2370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2371..2439,2440..3231)
                     /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(2589..2608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3402..3990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3891..3910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4144..4161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4234..4311
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4312..5394
                     /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(4331..4350)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5443..5448,5449..5466,5467..5473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5481..5497
                     /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     5505..5521
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5505..5521
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5517..6044
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5537..5554)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5537..5553)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5546..5568)
                     /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 cggatctggt tccgcgtgga tccctggaag ttctgttcca ggggcccatg
      781 gaacggctgc agaagcaacc acttacctcc cccgggagcg tgagcccctc ccgagattcc
      841 agtgtgcctg gctctccctc cagcatcgtg gccaagatgg acaatcaggt gctgggctac
      901 aaggacctgg ctgccatccc caaggacaag gccatcctgg acatcgagcg gcccgacctc
      961 atgatctacg agcctcactt cacttattcc ctcctggaac acgtggagct gcctcgcagc
     1021 cgcgagcgct cgctgtcacc caaagccgca gcggggggcg gagccgggaa ggtgtgggcg
     1081 gacagccggt cgcctggaat catctctcag gcctcggccc ccagaaccac tggaaccccc
     1141 cggaccagcc tgccccattt ccaccaccct gagacctccc gcccagattc caacatctac
     1201 aagaagcctc ccatctataa gcagagagag tccgtgggag gcagccctca gaccaagcac
     1261 ctcatcgagg atctcatcat cgagtcatcc aagtttcctg cagcccagcc cccagacccc
     1321 aaccagccag ccaaaatcga aaccgactac tggccatgcc ccccgtctct ggctgttgtg
     1381 gagacagagt ggaggaagcg gaaggcgtct cggaggggag cagaggaaga ggaggaggag
     1441 gaagatgacg actctggaga ggagatgaag gctctcaggg agcgtcagag agaggaactc
     1501 agtaaggtta cttccaactt gggaaagatg atcttgaaag aagagatgga aaagtcattg
     1561 ccgatccgaa ggaaaacccg ctctctgcct gaccggacac ccttccatac ctccttgcac
     1621 cagggaacgt ctaaatcttc ctctctcccc gcctatggca ggaccaccct gagccggcta
     1681 cagtccacag agttcagccc atcagggagt gagactggaa gcccaggcct gcagatctat
     1741 ccctatgaaa tgctagtggt gaccaacaag gggcgaacca agctgccacc gggggtggat
     1801 cggatgcggc ttgagaggca tctgtctgcc gaggacttct caagggtatt tgccatgtcc
     1861 cctgaagagt ttggcaagct ggctctgtgg aagcggaatg agctcaagaa gaaggcctct
     1921 ctcttccatc accatcacca tcactgatcc ccgggaattc atcgtgactg actgacgatc
     1981 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2041 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2101 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     2161 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     2221 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2281 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2341 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2401 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2461 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2521 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2581 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2641 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2701 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2761 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2821 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2881 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2941 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3001 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3061 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3121 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3181 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3241 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3301 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3361 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3421 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3481 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3541 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3601 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3661 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3721 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3781 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3841 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3901 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3961 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4021 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4081 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4141 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4201 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
     4261 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     4321 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     4381 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     4441 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     4501 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     4561 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     4621 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     4681 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     4741 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     4801 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     4861 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     4921 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     4981 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     5041 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     5101 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     5161 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     5221 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     5281 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5401 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     5461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     5521 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5581 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5641 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     5701 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     5761 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     5821 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     5881 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     5941 ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg
     6001 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     6061 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     6121 ctggcaaata ttctgaaatg agctg
//
