LOCUS       sequence-181748-        4967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of maize SAE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4967)
  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 4967)
  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
FEATURES             Location/Qualifiers
     source          1..4967
                     /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             1347..1391
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1429..1447)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1443..1490
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1711..2370)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2285..2304
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2371..2473)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2865..2882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2999..3544)
                     /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(3079..3098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3769..4851)
                     /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     4813..4832
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4852..4929)
                     /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 atggcagatc tcaattggat atcggccggc cacgcgatcg
     1321 ctgacgtcgg taccctcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc
     1381 acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat
     1441 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc
     1501 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc
     1561 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc
     1621 tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt
     1681 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta
     1741 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     1801 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa
     1861 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     1921 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     1981 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     2041 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     2101 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg
     2161 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     2221 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     2281 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     2341 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     2401 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     2461 tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
     2521 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
     2581 gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag
     2641 gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg
     2701 tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta
     2761 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     2821 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     2881 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     2941 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     3001 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     3061 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     3121 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     3181 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     3241 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3301 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     3361 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     3421 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     3481 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     3541 tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc aatttatctc
     3601 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     3661 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     3721 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3841 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     3901 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     3961 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4021 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     4081 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4141 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     4201 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     4261 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     4321 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     4381 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     4441 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     4501 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     4561 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     4621 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     4681 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     4741 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     4801 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     4861 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     4921 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcatt
//
