LOCUS       sequence-443734-        5465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ub with sortase motif and His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5465)
  AUTHORS   Antonenko A, Pomorski A, Singh AK, Kapczynska K, Krezel A
  TITLE     Site-Specific and Fluorescently Enhanced Installation of 
            Post-Translational Protein Modifications via Bifunctional 
            Biarsenical Linker.
  JOURNAL   ACS Omega. 2024 Oct 30;9(45):45127-45137. doi: 
            10.1021/acsomega.4c05828. eCollection 2024 Nov 12.
  PUBMED    39554417
REFERENCE   2  (bases 1 to 5465)
  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: 
            "10.1021/acsomega.4c05828"; journalName: "ACS Omega"; date: 
            "2024-10-30- 30"; volume: "9"; issue: "45"; pages: "45127-45137"
COMMENT     Sequence Label: Ub-LPNTG-pET
FEATURES             Location/Qualifiers
     source          1..5465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(734..1816)
                     /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     1778..1797
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1817..1894)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2100..2119
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2207..2226
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2207..2225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2226..2250
                     /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             2265..2287
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2532..2549
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2602..2620)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2616..2663
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2700..3155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2787..2806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2997..3018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3247..4062)
                     /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(3247..4062)
                     /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     3970..3989
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4054..4073
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4184..4772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4673..4692
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4926..4943
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4958..5097
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5023..5042)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5183..5205
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5199..5390)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
       61 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
      121 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
      181 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
      241 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
      301 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
      361 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
      421 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
      481 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
      541 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
      601 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
      661 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
      721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
      841 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
      901 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
      961 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     1021 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     1081 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     1141 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     1201 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     1261 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     1321 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     1381 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     1441 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     1501 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     1561 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     1621 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     1681 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     1741 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     1801 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     1861 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     1921 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     1981 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     2041 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     2101 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     2161 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact
     2221 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     2281 aaggagatat acatatgcag atcttcgtca agacgttaac cggtaaaacc ataactctgg
     2341 aagttgaacc atccgatacc atcgaaaacg ttaaggctaa aattcaagac aaggaaggaa
     2401 ttccacctga tcaacaaaga ttgatctttg ccggtaagca gctcgaggac ggtagaacgc
     2461 tgtctgatta caacattcag aaggagtcga ccttacatct tgtcttaaga ctaccgaaca
     2521 ccggtggatc ccaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     2581 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     2641 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     2701 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2761 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2821 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2881 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2941 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3001 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3061 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3121 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     3181 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     3241 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     3301 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     3361 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     3421 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     3481 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     3541 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     3601 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     3661 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     3721 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     3781 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3841 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3901 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     3961 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     4021 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     4081 gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt
     4141 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4201 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4261 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4321 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4381 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4441 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4501 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4561 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4621 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4681 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4741 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4801 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4861 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4921 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4981 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     5041 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     5101 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5161 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5221 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     5281 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     5341 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     5401 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     5461 acgat
//
