LOCUS       sequence-459092-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H43K SUMO1 protein under T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  AUTHORS   Kim J, Qin S, Zhou HX, Rosen MK
  TITLE     Surface Charge Can Modulate Phase Separation of Multidomain 
            Proteins.
  JOURNAL   J Am Chem Soc. 2024 Feb 7;146(5):3383-3395. doi: 
            10.1021/jacs.3c12789. Epub 2024 Jan 23.
  PUBMED    38262618
REFERENCE   2  (bases 1 to 5689)
  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/jacs.3c12789"; journalName: "J Am Chem Soc"; date: 
            "2024-02-7- 7"; volume: "146"; issue: "5"; pages: "3383-3395"
COMMENT     Sequence Label: H43K SUMO1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     982..1001
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1021..1098)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1304..1323
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1407..1426
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1407..1425
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1426..1450
                     /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             1465..1487
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1501..1524
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1525..1545
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1887..1905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1973..2077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2078..2146,2147..2938)
                     /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(2296..2315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3109..3697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3598..3617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3851..3868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3883..4022
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3948..3967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4108..4130
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4124..4315)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5627..5689,1..1020))
                     /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"
ORIGIN
        1 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
       61 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
      121 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
      181 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
      241 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
      301 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
      361 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
      421 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
      481 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
      541 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
      601 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
      661 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
      721 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
      781 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
      841 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
      901 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
      961 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     1021 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     1081 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     1141 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     1201 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     1261 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     1321 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     1381 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     1441 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat acatatcatg
     1501 caccaccatc accaccatca tcacgaaaac ctgtattttc agggcatgca tgaattcggc
     1561 ggctcatggg gtggttctat gagtgatcag gaagccaaac cctcaacgga ggatcttggt
     1621 gataaaaaag aaggggagta tattaaactg aaagtgattg gacaggacag tagcgagatt
     1681 catttcaaag ttaagatgac tacgaagctg aaaaagctga aggaatcgta ttgccagcgc
     1741 cagggtgttc cgatgaacag cttacgcttc ctgttcgaag gccagcgcat cgcggacaac
     1801 catactccta aagagctggg tatggaagag gaggacgtga ttgaagtata ccaggaacag
     1861 accggcggat aacaaagccc gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     1921 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1981 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2041 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2101 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2161 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2221 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2281 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     2341 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2401 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2461 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2521 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2581 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     2641 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2701 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2761 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2821 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2881 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2941 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3001 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3061 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3241 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3481 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3541 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3601 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3661 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3721 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3781 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3841 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3901 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     3961 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     4021 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4081 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4141 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4201 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4261 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     4321 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     4381 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     4441 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     4501 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     4561 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     4621 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     4681 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     4741 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     4801 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     4861 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     4921 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     4981 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     5041 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     5101 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     5161 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     5221 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     5281 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     5341 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     5401 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     5461 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     5521 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     5581 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     5641 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgc
//
