LOCUS       sequence-421138-        5975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H6-Ulp1 from pHYRS52 into pET-28b(+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5975)
  AUTHORS   Guerrero F, Ciragan A, Iwai H
  TITLE     Tandem SUMO fusion vectors for improving soluble protein expression 
            and purification.
  JOURNAL   Protein Expr Purif. 2015 Aug 20. pii: S1046-5928(15)30039-5. doi: 
            10.1016/j.pep.2015.08.019.
  PUBMED    26297996
REFERENCE   2  (bases 1 to 5975)
  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"; journalName: "Protein 
            Expr Purif. 2015 Aug 20. pii: S1046-5928(15)30039-5. doi: 
            10.1016/j.pep.2015.08.019."
COMMENT     Sequence Label: pFGET19_Ulp1
FEATURES             Location/Qualifiers
     source          1..5975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..44)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      40..87
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      124..579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(211..230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     421..442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(671..1486)
                     /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(671..1486)
                     /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     1394..1413
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1478..1497
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1608..2196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2097..2116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2350..2367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2382..2521
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2447..2466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2607..2629
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2623..2814)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3623..4705)
                     /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     4667..4686
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4706..4783)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4989..5008
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5092..5111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5092..5110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5111..5135
                     /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)."
     RBS             5150..5172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5192..5209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5213..5230
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5931..5948
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
       61 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
      121 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      181 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      241 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      301 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      361 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      421 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      481 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      541 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
      601 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      661 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
      721 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
      781 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
      841 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
      901 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
      961 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1021 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1081 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1141 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1201 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1261 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1321 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1381 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1441 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     1501 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
     1561 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1621 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1681 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1741 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1801 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1861 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1921 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1981 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2041 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2101 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2161 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2221 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2281 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2341 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     2401 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     2701 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     2761 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     2821 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     2881 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     2941 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     3001 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     3061 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     3121 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     3181 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     3241 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat
     3301 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     3361 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     3421 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     3481 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     3541 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3601 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3661 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3721 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3781 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3841 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3901 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3961 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4021 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4081 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4141 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     4201 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     4261 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4321 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     4381 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     4441 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4501 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4561 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4621 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4681 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4741 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     4801 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     4861 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     4921 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     4981 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     5041 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
     5101 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     5161 ttaagaagga gatatacata tgcggggttc tcatcatcat catcatcatg gtctggttcc
     5221 gcgtggatcc cttgttcctg aattaaatga aaaagacgat gaccaagtac aaaaagcttt
     5281 ggcatctaga gaaaatactc agttaatgaa tagagataat atagagataa cagtacgtga
     5341 ttttaagacc ttggcaccac gaagatggct aaatgacact atcattgagt tttttatgaa
     5401 atacattgaa aaatctaccc ctaatacagt ggcgtttaat tcgtttttct ataccaattt
     5461 atcagaaagg ggttatcaag gcgtccggag gtggatgaag agaaagaaga cacaaattga
     5521 taaacttgat aaaatcttta caccaataaa tttgaaccaa tcccactggg cgttgggcat
     5581 aattgattta aaaaagaaaa ctataggtta cgtagattca ttatcgaatg gtccaaatgc
     5641 tatgagtttc gctatactga ctgacttgca aaaatatgtt atggaggaaa gtaagcatac
     5701 aataggagaa gactttgatt tgattcattt agattgtccg cagcaaccaa atggctacga
     5761 ctgtggaata tatgtttgta tgaatactct ctatggaagt gcagatgcgc cattggattt
     5821 tgattataaa gatgcgatta ggatgagaag atttattgcc catttgattt taaccgacgc
     5881 tttaaaataa gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc
     5941 accaccactg agatccggct gctaacaaag cccga
//
