LOCUS       sequence-181741-        6434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of maize SAE1 and truncated SAE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6434)
  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 6434)
  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: pACYC_HA-ZmSAE1_ZmSAE2(TRUNC)-FLAG
FEATURES             Location/Qualifiers
     source          1..6434
                     /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             102..128
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1215..1239
                     /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             2770..2793
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2815..2859
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2897..2915)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2911..2958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3179..3838)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3753..3772
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3839..3941)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4333..4350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4467..5012)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4547..4566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5237..6319)
                     /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     6281..6300
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6320..6397)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taggaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctgtaga
       61 aataattttg tttaacttta ataaggagat ataccatggg ctacccgtat gatgtgccgg
      121 attatgcgat ggacggcggc ggcggcgctg gcggcgcgga gggggagctc acggcgcagg
      181 agacagcgct ctacgaccgc cagatccgcg tctggggcgt cgatgcccag aagagactaa
      241 gtaaggcgca cgtgcttgtg tgtggcatga acggaaccac catcgagttc tgcaagaata
      301 ttgttctagc tggagttgga agtttatcct tgatggatga taatgtagtc acagaggagg
      361 atctgaatgc aaatttccta attcctcctg atgagagcat atatggtggt agatctcgag
      421 cccaagtttg ctgcgagtct ttgatagatt tcaatcctat ggttcgagtt tttgttgaaa
      481 agggtgatcc atcattaatt gatggagaat tccttgacaa gtttgacatt gttgtactta
      541 gccgtgcatc tcttaaaaca aagttgttga ttaatgaaaa ctgtcgaaag agaagcaaac
      601 acattgcatt ttatactata gattgcaagg attcatgtgg tgaaatattt gttgatttac
      661 agaagcatag ctatgttcag aaaaagcctg gaggagcaac tgaacaacag gagttgacat
      721 attctagcct tcaggaagct atctctgtgc cttggaatag tctaccgaag aagacggcaa
      781 aattatactt tgctatgaga gtactggagg attatgaatt atctgaaggg cgcagccctg
      841 gcgaaacaac actttctgat atagatgcag ttctggctcg gaggaaggat atgtgtgata
      901 aaatgtcttt gaatgagtct cgtattccta ctactctcgt ggaacgactg ctagcagctg
      961 gaaagaagga gcatcctcct gtatgtgcta tccttggtgg tattcttggt caggaggtga
     1021 tcaaatcgat atcctgcaag ggcgatccag tcaagaattt cttctatttc gatgtggctg
     1081 acggtaaagg tgtcatcgag gatatcccac caccaccacc tgcaaaataa gcggccgcat
     1141 aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata
     1201 cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa
     1261 gtataagaag gagatataca tatggcagcc gtcgcgtcct cggaggaggc cgtcaaggca
     1321 tcgaaggtgc tgatggtcgg ggcgggaggc atcggctgcg agctgctcaa gacgctcgcg
     1381 ctctccggat tcagcgacat acacatcatt gacttggata caattgaagt tagcaatctg
     1441 aatagacagt ttttgtttcg gcagagtcat gttggtcagt caaaagctaa agttgctcgg
     1501 gatgctgtat tgaaatttag gccaaatatc aatataaacc cataccatgc aaacgtgaag
     1561 gattctcatt tcaatgttga tttcttcaag caatttaatg ttgttctgaa tggtcttgat
     1621 aacttggatg ctagacgcca tgttaatcgc ctttgccttg ctgctgatgt tcctttggtt
     1681 gaaagtggta ctactggttt tttgggacag gttactgttc atgtcaaggg taaaacagag
     1741 tgctatgagt gtcaaccaaa gcctgtccct aaatcatatc ctgtctgcac aattacaaac
     1801 acaccatcaa agtttgttca ttgcattgtt tgggcgaaag atctgctttt tgcaaaatta
     1861 tttggtgata aaaaacagga taatgatctt aatgtgcact ccaaagatga taccagttca
     1921 aaaacagatg tttttgaaag gagtgtagat gaagatcttg agcaatacac ccagagaata
     1981 tatgaccacg tatttggtta caacattgaa gttgcattgg ataataagga aacttggaag
     2041 aatcggagaa agccaaaccc tatttacatt agagatgcat tgcctgaaga cgctgttcaa
     2101 caaaatggtc gctctcagga tcacgtgaat gaggagcagg atccttctgc tatggtatcc
     2161 ttaggcctta gaaatacaca agagatatgg agtcttgctg acaattcaag agtgttcttg
     2221 gaggctttaa aattattttt tgacaaaaga gagaaggaaa tagggagcct tatttttgac
     2281 aaggacgacc agttggctgt tgagtttgtt accgctgcag caaatatcag agcttcctct
     2341 tttggaatac cactgcatag tctttttgaa gctaaaggtg ttgctggaaa tatagttcat
     2401 gctgtggcaa ctaccaatgc tataattgct ggtctaatcg tcatcgaggc aattaaagtg
     2461 ctcaagggtg attatcaaga ttatagaatg acatattgcc ttgagcatcc agcaaggaag
     2521 atgctcttga tgcctgtcga gccttttgaa cctagtaaat catgttatgt ctgctctgag
     2581 accccacttg ttctggaggt taataccaag acaacaaagc tccgggaagt cattgaaaag
     2641 gttataaaag gtaagcttgg aatgaaccta cctttgataa tggttggcgc cacacttgtt
     2701 tttgaggatg gtgaagatct ggaggaggat gagattgcaa attatgcttt aaaccttgaa
     2761 aaggtgctgg attataaaga tgatgatgat aaataaggta ccctcgagtc tggtaaagaa
     2821 accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta
     2881 acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2941 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt
     3001 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     3061 cctgaaccga cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat
     3121 cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct taaaaaaatt
     3181 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     3241 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     3301 cttgcgtata atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc atattggcca
     3361 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     3421 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     3481 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     3541 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     3601 cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa
     3661 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     3721 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     3781 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     3841 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     3901 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     3961 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     4021 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac
     4081 tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt
     4141 ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag
     4201 cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc
     4261 ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat
     4321 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga
     4381 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga
     4441 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac
     4501 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg
     4561 tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc
     4621 attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc
     4681 agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt
     4741 atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc
     4801 agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa
     4861 ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt
     4921 tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc
     4981 aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat
     5041 atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     5101 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     5161 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     5341 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     5401 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     5461 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     5521 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     5581 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5641 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5701 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5761 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5821 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5881 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     5941 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     6001 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     6061 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     6121 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     6181 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     6241 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     6301 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     6361 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     6421 tgcgactcct gcat
//
