LOCUS       sequence-181738-        4093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of maize SUMO1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4093)
  AUTHORS   Augustine RC, York SL, Rytz TC, Vierstra RD
  TITLE     Defining the SUMO System in Maize: SUMOylation Is Up-Regulated 
            during Endosperm Development and Rapidly Induced by Stress.
  JOURNAL   Plant Physiol. 2016 Jul;171(3):2191-210. doi: 10.1104/pp.16.00353. 
            Epub 2016 May 15.
  PUBMED    27208252
REFERENCE   2  (bases 1 to 4093)
  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: 
            "10.1104/pp.16.00353"; journalName: "Plant Physiol"; date: 
            "2016-07"; volume: "171"; issue: "3"; pages: "2191-210"
COMMENT     Sequence Label: pRSF_6His-ZmSUMO1a-GG
FEATURES             Location/Qualifiers
     source          1..4093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..27
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    28..52
                     /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)."
     CDS             108..125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        503..521
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    522..546
                     /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)."
     CDS             655..699
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(737..755)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      751..798
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1028..1843)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1028..1843)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1751..1770
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1844..1935)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1951..2700)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(1961..1978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2201..2220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2896..3978)
                     /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     3940..3959
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3979..4056)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taggaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctgtaga
       61 aataattttg tttaacttta ataaggagat ataccatggg cagcagccat caccatcatc
      121 accacagcca ggatccgaat tcgatgtctg gagccggcga ggaggataag aagcccgcgg
      181 agggcggcgc ccacatcaac cttaaggtca agggacagga tggcaatgag gtgttctttc
      241 gcataaagag gtccacccag ctgaagaagc tgatgaacgc ctattgcgac cgccagtctg
      301 tggacatgaa tgccattgca ttcctgtttg atggccgcag gcttcgcggc gagcagaccc
      361 ctgatgagct ggagatggag gacggcgacg agatcgacgc catgcttcac cagaccggag
      421 gctaggtcga caagcttgcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg
      481 tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac
      541 aattccccat cttagtatat tagttaagta taagaaggag atatacatat ggcagatctc
      601 aattggatat cggccggcca cgcgatcgct gacgtcggta ccctcgagtc tggtaaagaa
      661 accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta
      721 acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      781 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt
      841 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
      901 cctgaaccga cgacaagctg acgaccgggt ctccgcaagt ggcacttttc ggggaaatgt
      961 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     1021 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     1081 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     1141 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     1201 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     1261 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     1321 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     1381 gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag
     1441 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     1501 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     1561 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     1621 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     1681 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     1741 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     1801 atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat
     1861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     1921 agaaaaataa acaaataggc atgcagcgct cttccgcttc ctcgctcact gactcgctac
     1981 gctcggtcgt tcgactgcgg cgagcggtgt cagctcactc aaaagcggta atacggttat
     2041 ccacagaatc aggggataaa gccggaaaga acatgtgagc aaaaagcaaa gcaccggaag
     2101 aagccaacgc cgcaggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2161 cgacgctcaa gccagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgttg gtatctcagt
     2341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2461 ccactggcag cagccattgg taactgattt agaggacttt gtcttgaagt tatgcacctg
     2521 ttaaggctaa actgaaagaa cagattttgg tgagtgcggt cctccaaccc acttaccttg
     2581 gttcaaagag ttggtagctc agcgaacctt gagaaaacca ccgttggtag cggtggtttt
     2641 tctttattta tgagatgatg aatcaatcgg tctatcaagt caacgaacag ctattccgtt
     2701 actctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     2761 atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg
     2821 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     2881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     3001 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     3061 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     3121 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc
     3181 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     3241 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     3301 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     3361 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     3421 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     3481 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     3541 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     3601 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     3661 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     3721 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     3781 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     3841 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     3901 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     3961 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gatatcatgc
     4021 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     4081 gcgactcctg cat
//
