LOCUS       sequence-181727-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of maize SUMO1a and inactive SCE1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4552)
  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 4552)
  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_ZmSCE1b(C94S)-Myc
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..62
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    63..87
                     /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             143..160
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        538..556
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    557..581
                     /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             1104..1133
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1149..1193
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1231..1249)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1245..1292
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1522..2337)
                     /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(1522..2337)
                     /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     2245..2264
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2338..2429)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2445..3194)
                     /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(2455..2472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2695..2714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3390..4472)
                     /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     4434..4453
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4473..4550)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggatctcgac gctctccctt atgcgactcc tgcattagga aattaatacg actcactata
       61 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
      121 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgat
      181 gtctggagcc ggcgaggagg ataagaagcc cgcggagggc ggcgcccaca tcaaccttaa
      241 ggtcaaggga caggatggca atgaggtgtt ctttcgcata aagaggtcca cccagctgaa
      301 gaagctgatg aacgcctatt gcgaccgcca gtctgtggac atgaatgcca ttgcattcct
      361 gtttgatggc cgcaggcttc gcggcgagca gacccctgat gagctggaga tggaggacgg
      421 cgacgagatc gacgccatgc ttcaccagac cggaggctag gtcgacaagc ttgcggccgc
      481 ataatgctta agtcgaacag aaagtaatcg tattgtacac ggccgcataa tcgaaattaa
      541 tacgactcac tataggggaa ttgtgagcgg ataacaattc cccatcttag tatattagtt
      601 aagtataaga aggagatata catatgtctg ggggaatcgc ccgcggccgc cttgccgagg
      661 agcgcaaggc ctggcgcaag aaccacccgc acggtttcgt cgcaaagccg gagtcgctgc
      721 ccgacgggac ggtgaacctg atgatctggc agtgcaccat ccccggcaag caagggactg
      781 actgggaagg tggatatttc cctctcaccc ttcattttag tgaggattac cctagcaagc
      841 ctcccaagtg caagttccct cagggtttct tccacccaaa tgtgtatcct tctggaacag
      901 tctccctttc gattcttaat gaggacagtg gttggagacc agctattact gttaagcaga
      961 ttctcgtcgg gatccaggac ttgctagatc agccaaatcc tgctgaccct gctcaaacag
     1021 acggatatca cctttttatc caggatccca cagaatacaa gaggcgcgtt agactgcagg
     1081 cgaagcagta tcctgcattg gtcgaacaaa aactcatctc agaagaggat ctgtgactcg
     1141 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     1201 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     1261 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
     1321 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     1381 tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact
     1441 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1501 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1561 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1621 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1681 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1741 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1801 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1861 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg
     1921 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1981 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     2041 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2101 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2161 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2221 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2281 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact
     2341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2401 atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct
     2461 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc
     2521 ggtaatacgg ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag
     2581 caaagcaccg gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg
     2641 agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat
     2701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2821 gttggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2941 gacacgactt atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg
     3001 aagttatgca cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca
     3061 acccacttac cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg
     3121 gtagcggtgg tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga
     3181 acagctattc cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
     3241 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
     3301 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     3361 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3421 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3481 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     3541 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     3601 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     3661 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     3721 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     3781 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     3841 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     3901 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     3961 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     4021 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     4081 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     4141 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     4201 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     4261 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     4321 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     4381 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     4441 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     4501 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cg
//
