LOCUS       sequence-220213-        6127 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  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 6127)
  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
FEATURES             Location/Qualifiers
     source          1..6127
                     /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"
     primer_bind     complement(1995..2017)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2162..2180)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2248..2352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2353..2421,2422..3213)
                     /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(2571..2590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3384..3972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3873..3892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4126..4143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4216..4293
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4294..5376
                     /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(4313..4332)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5425..5430,5431..5448,5449..5455)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5463..5479
                     /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     5487..5503
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5487..5503
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5499..6026
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5519..5536)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5519..5535)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5528..5550)
                     /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 ctcttctgat ccccgggaat tcatcgtgac tgactgacga tctgcctcgc gcgtttcggt
     1981 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2041 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2101 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt ataattcttg aagacgaaag
     2161 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2221 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2341 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2401 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2461 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2521 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2581 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2641 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2701 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2761 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2821 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2881 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2941 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3001 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3061 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3121 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3181 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3241 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3301 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3361 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3421 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3481 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3541 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3601 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3661 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3721 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3781 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3841 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3901 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3961 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4021 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4081 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4141 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4201 caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     4261 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     4321 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     4381 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     4441 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     4501 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     4561 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     4621 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     4681 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     4741 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     4801 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     4861 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     4921 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     4981 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     5041 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     5101 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     5161 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     5221 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     5281 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     5341 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     5401 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     5461 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac
     5521 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5581 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     5641 cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag
     5701 aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc
     5761 cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca
     5821 ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat
     5881 ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg
     5941 gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag
     6001 ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact
     6061 cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa
     6121 tgagctg
//
