LOCUS       sequence-181744-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of maize SAE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  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 5903)
  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_ZmSAE2-FLAG
FEATURES             Location/Qualifiers
     source          1..5903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..35
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    36..60
                     /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             116..133
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        247..265
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    266..290
                     /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             2247..2270
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2292..2336
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2374..2392)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2388..2435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2656..3315)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3230..3249
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3316..3418)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3944..4489)
                     /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(4024..4043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4714..5796)
                     /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     5758..5777
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5797..5874)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
       61 cctgtagaaa taattttgtt taactttaat aaggagatat accatgggca gcagccatca
      121 ccatcatcac cacagccagg atccgaattc gagctcggcg cgcctgcagg tcgacaagct
      181 tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
      241 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
      301 atattagtta agtataagaa ggagatatac atatggcagc cgtcgcgtcc tcggaggagg
      361 ccgtcaaggc atcgaaggtg ctgatggtcg gggcgggagg catcggctgc gagctgctca
      421 agacgctcgc gctctccgga ttcagcgaca tacacatcat tgacttggat acaattgaag
      481 ttagcaatct gaatagacag tttttgtttc ggcagagtca tgttggtcag tcaaaagcta
      541 aagttgctcg ggatgctgta ttgaaattta ggccaaatat caatataaac ccataccatg
      601 caaacgtgaa ggattctcat ttcaatgttg atttcttcaa gcaatttaat gttgttctga
      661 atggtcttga taacttggat gctagacgcc atgttaatcg cctttgcctt gctgctgatg
      721 ttcctttggt tgaaagtggt actactggtt ttttgggaca ggttactgtt catgtcaagg
      781 gtaaaacaga gtgctatgag tgtcaaccaa agcctgtccc taaatcatat cctgtctgca
      841 caattacaaa cacaccatca aagtttgttc attgcattgt ttgggcgaaa gatctgcttt
      901 ttgcaaaatt atttggtgat aaaaaacagg ataatgatct taatgtgcac tccaaagatg
      961 ataccagttc aaaaacagat gtttttgaaa ggagtgtaga tgaagatctt gagcaataca
     1021 cccagagaat atatgaccac gtatttggtt acaacattga agttgcattg gataataagg
     1081 aaacttggaa gaatcggaga aagccaaacc ctatttacat tagagatgca ttgcctgaag
     1141 acgctgttca acaaaatggt cgctctcagg atcacgtgaa tgaggagcag gatccttctg
     1201 ctatggtatc cttaggcctt agaaatacac aagagatatg gagtcttgct gacaattcaa
     1261 gagtgttctt ggaggcttta aaattatttt ttgacaaaag agagaaggaa atagggagcc
     1321 ttatttttga caaggacgac cagttggctg ttgagtttgt taccgctgca gcaaatatca
     1381 gagcttcctc ttttggaata ccactgcata gtctttttga agctaaaggt gttgctggaa
     1441 atatagttca tgctgtggca actaccaatg ctataattgc tggtctaatc gtcatcgagg
     1501 caattaaagt gctcaagggt gattatcaag attatagaat gacatattgc cttgagcatc
     1561 cagcaaggaa gatgctcttg atgcctgtcg agccttttga acctagtaaa tcatgttatg
     1621 tctgctctga gaccccactt gttctggagg ttaataccaa gacaacaaag ctccgggaag
     1681 tcattgaaaa ggttataaaa ggtaagcttg gaatgaacct acctttgata atggttggcg
     1741 ccacacttgt ttttgaggat ggtgaagatc tggaggagga tgagattgca aattatgctt
     1801 taaaccttga aaaggtcttg gctgaacttc cagctccagt tctaaatggt acaatggtta
     1861 cagtcgagga tttgcatcag gaactaaaat gcagtatcaa catcaaacac agggatgagt
     1921 tcgatgaaga aaaggagcct gacagaatgg ttcttgttgg gtggtctggc ccagttgata
     1981 agcaaaccac tagtaatggt gagactaaat cagttccatc ttcatccaat acagaagatg
     2041 tggatggtac tgccgaggac atatcagcaa cttctggaat gaagcgtaag ctgaacgaaa
     2101 tattagagac aaaggagaat tttgatgctc tcgaaaatct ttcgggtgtt gattccagca
     2161 gcgctcaagt tgttgaagat gacaatgacg atggccttct tatgtttgat gaggacccaa
     2221 ggcaaagtaa gaagaagaga ttgcaggatt ataaagatga tgatgataaa taaggtaccc
     2281 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     2341 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     2401 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     2461 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     2521 ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc
     2581 attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata
     2641 actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     2701 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     2761 catcagcacc ttgtcgcctt gcgtataata tttgcccata gtgaaaacgg gggcgaagaa
     2821 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     2881 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     2941 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     3001 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     3061 ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag cattcatcag
     3121 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     3181 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     3241 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     3301 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     3361 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     3421 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     3481 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
     3541 tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt
     3601 ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa
     3661 gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
     3721 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     3781 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     3841 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     3901 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     3961 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     4021 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     4081 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     4141 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     4201 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     4261 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     4321 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     4381 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     4441 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     4501 tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc
     4561 tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac
     4621 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4681 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4741 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4801 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4861 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4921 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4981 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     5041 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     5101 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     5161 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     5221 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5281 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     5341 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     5401 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     5461 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     5521 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5581 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5641 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5701 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5761 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5821 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5881 ctcgacgctc tcccttatgc gac
//
