LOCUS       sequence-193567-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains HCF-1rep1wt with GST-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  AUTHORS   Capotosti F, Guernier S, Lammers F, Waridel P, Cai Y, Jin J, Conaway
            JW, Conaway RC, Herr W
  TITLE     O-GlcNAc transferase catalyzes site-specific proteolysis of HCF-1.
  JOURNAL   Cell. 2011 Feb 4;144(3):376-88. doi: 10.1016/j.cell.2010.12.030.
  PUBMED    21295698
REFERENCE   2  (bases 1 to 5615)
  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.cell.2010.12"; journalName: "Cell"; date: "2011-02-4- 4";
            volume: "144"; issue: "3"; pages: "376-88"
COMMENT     Sequence Label: pGEX-HCF-1rep1wt
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(188..193,194..209,210..216)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    224..240
                     /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             263..916
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     874..896
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             923..946
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             950..970
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1604..1621
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1670..1692)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1837..1855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1923..2027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2028..2096,2097..2888)
                     /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(2246..2265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3059..3647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3548..3567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3801..3818
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3891..3968
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3969..5051
                     /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(3988..4007)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5100..5105,5106..5123,5124..5130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5154
                     /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     5162..5178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5162..5178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5174..5615,1..86)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5194..5211)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5194..5210)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5203..5225)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag
       61 ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact
      121 cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa
      181 tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa
      241 tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct
      301 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
      361 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
      421 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
      481 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
      541 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
      601 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
      661 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
      721 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
      781 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
      841 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
      901 cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggcccctgg aaaacttgta
      961 tttccagggc gggtccccag ccgtcaccac gttggttgtg aaaggcacca caggtgtcac
     1021 gaccctaggc acagtgacag gcaccgtctc caccagcctt gccggggcgg ggggccacag
     1081 cactagtgct tccctggcca cgcccatcac caccttgggc accattgcca ccctctcaag
     1141 ccaggtgatc aaccccactg ccatcactgt gtcggccgca cagaccacgc tgacagcggc
     1201 aggcgggctc acaaccccaa ccatcaccat gcagcccgtg tcccagccca cccaggtaac
     1261 tctgatcacg gcacctagtg gggtggaggc ccagcctgtg catgacctcc ctgtgtccat
     1321 tctggcctcc ccgactacag aacagcccac cgccacagtt accatcgccg actcaggcca
     1381 gggtgatgtg cagcctggca ctgtcacctt ggtgtgctcc aacccaccct gtgagaccca
     1441 cgagactggc accaccaaca cggccaccac tactgttgtg gctaaccttg ggggacaccc
     1501 ccagcccacc caagtgcagt tcgtctgtga cagacaggag gcagctgctt ctcttgtgac
     1561 ctcgactgtg ggccagcaga atggtagcgt ggcggccgca tcgcaccatc atcatcatca
     1621 ttgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     1801 gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     1861 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1921 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1981 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2041 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2101 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2161 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2221 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2281 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2341 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2401 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2461 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2521 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     2581 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2641 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2701 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2761 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2821 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2881 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2941 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3001 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3121 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3181 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3541 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3601 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3661 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3721 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3781 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3841 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg
     3901 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     3961 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     4021 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     4081 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     4141 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     4201 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     4261 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     4321 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     4381 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     4441 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     4501 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     4561 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     4621 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     4681 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     4741 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     4801 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     4861 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     4921 tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct
     4981 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5041 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
     5101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5161 acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga
     5221 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5281 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5341 tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga
     5401 gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta
     5461 cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc
     5521 cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca
     5581 ggaaggccag acgcgaatta tttttgatgg cgttg
//
