LOCUS       sequence-459218-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of E67K 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: E67K SUMO1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             472..663
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(657..679)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    765..904
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     820..839
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(919..936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1090..1678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1170..1189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1849..2640,2641..2709))
                     /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     2472..2491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2710..2814)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2882..2900
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(3242..3262)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3263..3286)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             3300..3322
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3337..3361
                     /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(3361..3380)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3362..3380)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3464..3483)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3689..3766
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3767..4849
                     /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(3786..3805)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
       61 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
      121 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
      181 agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga
      241 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc
      301 tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt
      361 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
      421 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
      481 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
      541 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
      601 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
      661 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
      721 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
      781 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
      841 tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      901 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1501 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     1561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2161 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2461 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2821 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2881 aaataggcgt atcacgaggc cctttcgggc tttgttatcc gccggtctgt tcctggtata
     2941 cttcaatcac gtcctcctct tccataccca gctctttagg agtatggttg tccgcgatgc
     3001 gctggccttt gaacaggaag cgtaagctgt tcatcggaac accctggcgc tggcaatacg
     3061 attccttcag ctttttcaga tgcgtagtca tcttaacttt gaaatgaatc tcgctactgt
     3121 cctgtccaat cactttcagt ttaatatact ccccttcttt tttatcacca agatcctccg
     3181 ttgagggttt ggcttcctga tcactcatag aaccacccca tgagccgccg aattcatgca
     3241 tgccctgaaa atacaggttt tcgtgatgat ggtggtgatg gtggtgcatg atatgtatat
     3301 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
     3361 ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc
     3421 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     3481 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     3541 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     3601 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     3661 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
     3721 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     3781 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     3841 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     3901 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     3961 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     4021 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     4081 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     4141 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     4201 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     4261 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     4321 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     4381 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     4441 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     4501 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     4561 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     4621 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     4681 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     4741 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4801 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4861 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     4921 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     4981 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     5041 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     5101 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     5161 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
     5221 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
     5281 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
     5341 aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg
     5401 cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc
     5461 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     5521 gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca
     5581 attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc
     5641 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcc
//
