LOCUS       sequence-459094-        5986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of small ubiquitin-like modifier 3 (SUMO3) protein under 
            T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5986)
  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 5986)
  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: small ubiquitin-like modifier 3 (SUMO3)
FEATURES             Location/Qualifiers
     source          1..5986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             555..746
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(740..762)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    848..987
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     903..922
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1002..1019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1173..1761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1253..1272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1932..2723,2724..2792))
                     /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     2555..2574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2793..2897)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2965..2983
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(3010..3015,3016..3032,3033..3038)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(3213..3260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3256..3274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3622..3642)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3643..3666)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             3680..3702
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3717..3741
                     /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(3741..3760)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3742..3760)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3844..3863)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4069..4146
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4147..5229
                     /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(4166..4185)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc
       61 gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga
      121 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
      181 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
      241 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
      301 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
      361 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
      421 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
      481 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
      541 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
      601 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
      661 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
      721 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
      781 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      841 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
      901 tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga
      961 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     1021 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1081 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1141 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1201 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1261 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1321 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1381 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1441 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1501 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1561 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     1621 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1681 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2221 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     2281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2521 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     2581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2941 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     3001 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
     3061 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
     3121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     3181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
     3241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
     3301 cgggctttgt tatccgccgg tctgctgctg aaatacgtcg atagtgtctt catcctccat
     3361 ctccagctgt gccggcgtgt ccgtttcatt aatcggttga ccatcaaagc ggaagcgaat
     3421 ctgacgcata gaaagaccct ggcgttcgct gtacgctttc atcagtttcg acaacggtgt
     3481 gtggcgttta attttgaact gaaccacgga cccatcctgt ccagcaacct tcaagttaat
     3541 gtggtcattc tcggttttca cgccctcctt aggcttctct tctgacatag accctcccca
     3601 actcccaccc atatgatgca tgccctgaaa atacaggttt tcgtgatgat ggtggtgatg
     3661 gtggtgcatg atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
     3721 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct
     3781 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     3841 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     3901 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     3961 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
     4021 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
     4081 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     4141 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     4201 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     4261 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     4321 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     4381 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     4441 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     4501 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     4561 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     4621 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     4681 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     4741 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     4801 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     4861 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     4921 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     4981 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     5041 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     5101 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     5161 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5221 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
     5281 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     5341 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     5401 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     5461 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     5521 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
     5581 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
     5641 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
     5701 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat
     5761 tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc
     5821 atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
     5881 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact
     5941 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgc
//
