LOCUS       sequence-181746-        6860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of maize SAE1 and SAE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6860)
  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 6860)
  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-FLAG
FEATURES             Location/Qualifiers
     source          1..6860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..26
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    27..51
                     /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             101..127
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        1195..1213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1214..1238
                     /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             3195..3218
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3240..3284
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3322..3340)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3336..3383
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3604..4263)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4178..4197
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4264..4366)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4758..4775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4892..5437)
                     /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(4972..4991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5662..6744)
                     /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     6706..6725
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6745..6822)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aggaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctgtagaa
       61 ataattttgt ttaactttaa taaggagata taccatgggc tacccgtatg atgtgccgga
      121 ttatgcgatg gacggcggcg gcggcgctgg cggcgcggag ggggagctca cggcgcagga
      181 gacagcgctc tacgaccgcc agatccgcgt ctggggcgtc gatgcccaga agagactaag
      241 taaggcgcac gtgcttgtgt gtggcatgaa cggaaccacc atcgagttct gcaagaatat
      301 tgttctagct ggagttggaa gtttatcctt gatggatgat aatgtagtca cagaggagga
      361 tctgaatgca aatttcctaa ttcctcctga tgagagcata tatggtggta gatctcgagc
      421 ccaagtttgc tgcgagtctt tgatagattt caatcctatg gttcgagttt ttgttgaaaa
      481 gggtgatcca tcattaattg atggagaatt ccttgacaag tttgacattg ttgtacttag
      541 ccgtgcatct cttaaaacaa agttgttgat taatgaaaac tgtcgaaaga gaagcaaaca
      601 cattgcattt tatactatag attgcaagga ttcatgtggt gaaatatttg ttgatttaca
      661 gaagcatagc tatgttcaga aaaagcctgg aggagcaact gaacaacagg agttgacata
      721 ttctagcctt caggaagcta tctctgtgcc ttggaatagt ctaccgaaga agacggcaaa
      781 attatacttt gctatgagag tactggagga ttatgaatta tctgaagggc gcagccctgg
      841 cgaaacaaca ctttctgata tagatgcagt tctggctcgg aggaaggata tgtgtgataa
      901 aatgtctttg aatgagtctc gtattcctac tactctcgtg gaacgactgc tagcagctgg
      961 aaagaaggag catcctcctg tatgtgctat ccttggtggt attcttggtc aggaggtgat
     1021 caaatcgata tcctgcaagg gcgatccagt caagaatttc ttctatttcg atgtggctga
     1081 cggtaaaggt gtcatcgagg atatcccacc accaccacct gcaaaataag cggccgcata
     1141 atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg aaattaatac
     1201 gactcactat aggggaattg tgagcggata acaattcccc atcttagtat attagttaag
     1261 tataagaagg agatatacat atggcagccg tcgcgtcctc ggaggaggcc gtcaaggcat
     1321 cgaaggtgct gatggtcggg gcgggaggca tcggctgcga gctgctcaag acgctcgcgc
     1381 tctccggatt cagcgacata cacatcattg acttggatac aattgaagtt agcaatctga
     1441 atagacagtt tttgtttcgg cagagtcatg ttggtcagtc aaaagctaaa gttgctcggg
     1501 atgctgtatt gaaatttagg ccaaatatca atataaaccc ataccatgca aacgtgaagg
     1561 attctcattt caatgttgat ttcttcaagc aatttaatgt tgttctgaat ggtcttgata
     1621 acttggatgc tagacgccat gttaatcgcc tttgccttgc tgctgatgtt cctttggttg
     1681 aaagtggtac tactggtttt ttgggacagg ttactgttca tgtcaagggt aaaacagagt
     1741 gctatgagtg tcaaccaaag cctgtcccta aatcatatcc tgtctgcaca attacaaaca
     1801 caccatcaaa gtttgttcat tgcattgttt gggcgaaaga tctgcttttt gcaaaattat
     1861 ttggtgataa aaaacaggat aatgatctta atgtgcactc caaagatgat accagttcaa
     1921 aaacagatgt ttttgaaagg agtgtagatg aagatcttga gcaatacacc cagagaatat
     1981 atgaccacgt atttggttac aacattgaag ttgcattgga taataaggaa acttggaaga
     2041 atcggagaaa gccaaaccct atttacatta gagatgcatt gcctgaagac gctgttcaac
     2101 aaaatggtcg ctctcaggat cacgtgaatg aggagcagga tccttctgct atggtatcct
     2161 taggccttag aaatacacaa gagatatgga gtcttgctga caattcaaga gtgttcttgg
     2221 aggctttaaa attatttttt gacaaaagag agaaggaaat agggagcctt atttttgaca
     2281 aggacgacca gttggctgtt gagtttgtta ccgctgcagc aaatatcaga gcttcctctt
     2341 ttggaatacc actgcatagt ctttttgaag ctaaaggtgt tgctggaaat atagttcatg
     2401 ctgtggcaac taccaatgct ataattgctg gtctaatcgt catcgaggca attaaagtgc
     2461 tcaagggtga ttatcaagat tatagaatga catattgcct tgagcatcca gcaaggaaga
     2521 tgctcttgat gcctgtcgag ccttttgaac ctagtaaatc atgttatgtc tgctctgaga
     2581 ccccacttgt tctggaggtt aataccaaga caacaaagct ccgggaagtc attgaaaagg
     2641 ttataaaagg taagcttgga atgaacctac ctttgataat ggttggcgcc acacttgttt
     2701 ttgaggatgg tgaagatctg gaggaggatg agattgcaaa ttatgcttta aaccttgaaa
     2761 aggtcttggc tgaacttcca gctccagttc taaatggtac aatggttaca gtcgaggatt
     2821 tgcatcagga actaaaatgc agtatcaaca tcaaacacag ggatgagttc gatgaagaaa
     2881 aggagcctga cagaatggtt cttgttgggt ggtctggccc agttgataag caaaccacta
     2941 gtaatggtga gactaaatca gttccatctt catccaatac agaagatgtg gatggtactg
     3001 ccgaggacat atcagcaact tctggaatga agcgtaagct gaacgaaata ttagagacaa
     3061 aggagaattt tgatgctctc gaaaatcttt cgggtgttga ttccagcagc gctcaagttg
     3121 ttgaagatga caatgacgat ggccttctta tgtttgatga ggacccaagg caaagtaaga
     3181 agaagagatt gcaggattat aaagatgatg atgataaata aggtaccctc gagtctggta
     3241 aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt
     3301 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3361 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     3421 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     3481 cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt
     3541 attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa
     3601 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     3661 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     3721 gtcgccttgc gtataatatt tgcccatagt gaaaacgggg gcgaagaagt tgtccatatt
     3781 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     3841 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     3901 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     3961 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     4021 ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat
     4081 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     4141 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     4201 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     4261 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     4321 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     4381 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     4441 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     4501 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     4561 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     4621 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     4681 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     4741 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     4801 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     4861 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     4921 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     4981 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     5041 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     5101 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     5161 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     5221 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     5281 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     5341 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     5401 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     5461 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     5521 catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg
     5581 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     5641 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5701 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     5761 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     5821 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     5881 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     5941 gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     6001 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     6061 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     6121 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     6181 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     6241 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     6301 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     6361 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     6421 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     6481 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6541 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6601 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6661 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     6721 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     6781 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     6841 ccttatgcga ctcctgcatt
//
