LOCUS       sequence-459216-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H35K 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: H35K SUMO1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2288..2479
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2473..2495)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2581..2720
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2636..2655
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2735..2752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2906..3494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2986..3005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3665..4456,4457..4525))
                     /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     4288..4307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4526..4630)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4698..4716
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5058..5078)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5079..5102)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             5116..5138
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5153..5177
                     /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(5177..5196)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5178..5196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5280..5299)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5505..5582
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5583..5689,1..976)
                     /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(5602..5621)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
       61 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
      121 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
      181 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
      241 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
      301 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
      361 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
      421 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
      481 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
      541 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
      601 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
      661 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
      721 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
      781 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
      841 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
      901 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
      961 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat tgggcaccgg
     1021 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     1081 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     1141 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     1201 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     1261 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     1321 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     1381 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     1441 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     1501 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     1561 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     1621 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     1681 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     1741 cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
     1801 gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     1861 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     1921 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     1981 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     2041 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     2101 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     2161 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     2221 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     2281 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     2341 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     2401 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     2461 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2521 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2581 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     2641 tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aatatcgcac agatgcgtaa
     2701 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2761 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3481 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3661 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3721 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3781 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3841 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3901 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3961 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4021 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4081 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4141 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4201 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4261 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     4321 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4381 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4441 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4501 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4561 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4621 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4681 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgggctttg ttatccgccg
     4741 gtctgttcct ggtatacttc aatcacgtcc tcctcttcca tacccagctc tttaggagta
     4801 tggttgtccg cgatgcgctg gccttcgaac aggaagcgta agctgttcat cggaacaccc
     4861 tggcgctggc aatacgattc cttcagcttt ttcagatgcg tagtcatctt aactttgaat
     4921 ttaatctcgc tactgtcctg tccaatcact ttcagtttaa tatactcccc ttctttttta
     4981 tcaccaagat cctccgttga gggtttggct tcctgatcac tcatagaacc accccatgag
     5041 ccgccgaatt catgcatgcc ctgaaaatac aggttttcgt gatgatggtg gtgatggtgg
     5101 tgcatgatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
     5161 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat
     5221 cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc
     5281 ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc
     5341 ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc
     5401 cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg
     5461 cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc
     5521 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     5581 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     5641 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcggga
//
