LOCUS       sequence-459098-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H43R 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: H43R SUMO1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(102..122)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(123..146)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             160..182
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    197..221
                     /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     complement(221..240)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(222..240)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(324..343)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        549..626
                     /gene="lacI"
                     /label=lacI promoter
     CDS             627..1709
                     /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(646..665)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3021..3212
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3206..3228)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3314..3453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3369..3388
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3468..3485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3639..4227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3719..3738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4398..5189,5190..5258))
                     /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     5021..5040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5259..5363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5431..5449
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttaatatact ccccttcttt tttatcacca agatcctccg ttgagggttt ggcttcctga
       61 tcactcatag aaccacccca tgagccgccg aattcatgca tgccctgaaa atacaggttt
      121 tcgtgatgat ggtggtgatg gtggtgcatg atatgtatat ctccttctta aagttaaaca
      181 aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta
      241 atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg
      301 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
      361 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
      421 cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa
      481 cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg
      541 agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg
      601 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
      661 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
      721 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
      781 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
      841 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
      901 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
      961 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     1021 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     1081 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     1141 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     1201 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     1261 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     1321 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     1381 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     1441 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     1501 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     1561 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     1621 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1681 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact
     1741 cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     1801 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     1861 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     1921 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     1981 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     2041 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     2101 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     2161 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     2221 accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     2281 agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc
     2341 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     2401 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     2461 atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca
     2521 cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
     2581 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     2641 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     2701 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     2761 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     2821 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     2881 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     2941 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     3001 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     3061 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     3121 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     3181 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3241 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3301 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     3361 taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg
     3421 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     3481 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3541 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3601 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3661 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3721 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3781 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3841 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3901 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3961 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4021 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     4081 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4141 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4201 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4261 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4321 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4381 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4441 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4501 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4561 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4621 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4681 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     4741 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4801 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4861 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4921 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4981 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     5041 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5101 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5161 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5221 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5281 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5341 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5401 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgggct
     5461 ttgttatccg ccggtctgtt cctggtatac ttcaatcacg tcctcctctt ccatacccag
     5521 ctctttagga gtatggttgt ccgcgatgcg ctggccttcg aacaggaagc gtaagctgtt
     5581 catcggaaca ccctggcgct ggcaatacga ttccttcagc tttttcagac gcgtagtcat
     5641 cttaactttg aaatgaatct cgctactgtc ctgtccaatc actttcagt
//
