LOCUS       sequence-459087-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H75S 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: H75S SUMO1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             410..601
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(595..617)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    703..842
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     758..777
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(857..874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1028..1616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1108..1127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1787..2578,2579..2647))
                     /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     2410..2429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2648..2752)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2820..2838
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(3180..3200)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3201..3224)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             3238..3260
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3275..3299
                     /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(3299..3318)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3300..3318)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3402..3421)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3627..3704
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3705..4787
                     /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(3724..3743)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
       61 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
      121 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
      181 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
      241 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
      301 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
      361 cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca
      421 ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga
      481 gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca
      541 cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
      601 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
      661 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
      721 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
      781 agagtgcacc attgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      841 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      901 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      961 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1021 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1081 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1141 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1201 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1261 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1321 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1381 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1441 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     1501 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1561 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1621 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1681 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1741 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1801 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1861 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1921 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1981 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2041 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2101 ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2161 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2221 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2281 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2341 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2401 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2461 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2521 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2581 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2641 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2701 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2761 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     2821 ataggcgtat cacgaggccc tttcgggctt tgttatccgc cggtctgttc ctggtatact
     2881 tcaatcacgt cctcctcttc catacccagc tctttaggag tagagttgtc cgcgatgcgc
     2941 tggccttcga acaggaagcg taagctgttc atcggaacac cctggcgctg gcaatacgat
     3001 tccttcagct ttttcagatg cgtagtcatc ttaactttga aatgaatctc gctactgtcc
     3061 tgtccaatca ctttcagttt aatatactcc ccttcttttt tatcaccaag atcctccgtt
     3121 gagggtttgg cttcctgatc actcatagaa ccaccccatg agccgccgaa ttcatgcatg
     3181 ccctgaaaat acaggttttc gtgatgatgg tggtgatggt ggtgcatgat atgtatatct
     3241 ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc
     3301 ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat
     3361 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     3421 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     3481 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     3541 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     3601 gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta
     3661 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     3721 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     3781 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     3841 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     3901 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     3961 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     4021 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     4081 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     4141 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     4201 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     4261 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     4321 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     4381 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4441 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4501 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     4561 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4621 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     4681 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4741 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4801 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
     4861 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     4921 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
     4981 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     5041 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
     5101 caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg
     5161 acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg
     5221 cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa
     5281 ggatcgctcg cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg
     5341 atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta
     5401 taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga
     5461 atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat
     5521 tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg
     5581 ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc
     5641 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttg
//
