LOCUS       sequence-258135-        6837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  R over-expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6837)
  AUTHORS   Young JK, Gasior SL, Jones S, Wang L, Navarro P, Vickroy B, 
            Barrangou R
  TITLE     The repurposing of type I-E CRISPR-Cascade for gene activation in 
            plants.
  JOURNAL   Commun Biol. 2019 Oct 18;2:383. doi: 10.1038/s42003-019-0637-6. 
            eCollection 2019.
  PUBMED    31646186
REFERENCE   2  (bases 1 to 6837)
  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"; journalName: "Commun 
            Biol."; date: "2019-10-18"; volume: "2:383. doi"; pages: " 
            10.1038/s42003-019-0637-6. eCollection 2019"
COMMENT     Sequence Label: PHP11004
FEATURES             Location/Qualifiers
     source          1..6837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1238..1255)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1238..1254)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1247..1269)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1463..1482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1582..1604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1642..1660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1728..1832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1833..1901,1902..2693)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2051..2070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2864..3452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3353..3372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3606..3623
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3740..3761
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3776..3781,3782..3799,3800..3806)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3814..3830
                     /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)."
     primer_bind     3819..3841
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3838..3854
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3838..3854
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(6058..6837,1..1212))
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
ORIGIN
        1 ggctagagcc atcccaggat tccccaaaga gaaacactgg caagttagca atcagaacgt
       61 gtctgacgta caggtcgcat ccgtgtacga acgctagcag cacggatcta acacaaacac
      121 ggatctaaca caaacatgaa cagaagtaga actaccgggc cctaaccatg gaccggaacg
      181 ccgatctaga gaaggtagag aggggggggg gggaggacga gcggcgtacc ttgaagcgga
      241 ggtgccgacg ggtggatttg ggggagatct ggttgtgtgt gtgtgcgctc cgaacaacac
      301 gaggttgggg aaagagggtg tggagggggt gtctatttat tacggcgggc gaggaaggga
      361 aagcgaagga gcggtgggaa aggaatcccc cgtagctgcc ggtgccgtga gaggaggagg
      421 aggccgcctg ccgtgccggc tcacgtctgc cgctccgcca cgcaatttct ggatgccgac
      481 agcggagcaa gtccaacggt ggagcggaac tctcgagagg ggtccagagg cagcgacaga
      541 gatgccgtgc cgtctgcttc gcttggcccg acgcgacgct gctggttcgc tggttggtgt
      601 ccgttagact cgtcgacggc gtttaacagg ctggcattat ctactcgaaa caagaaaaat
      661 gtttccttag tttttttaat ttcttaaagg gtatttgttt aatttttagt cactttattt
      721 tattctattt tatatctaaa ttattaaata aaaaaactaa aatagagttt tagttttctt
      781 aatttagagg ctaaaataga ataaaataga tgtactaaaa aaattagtct ataaaaacca
      841 ttaaccctaa accctaaatg gatgtactaa taaaatggat gaagtattat ataggtgaag
      901 ctatttgcaa aaaaaaagga gaacacatgc acactaaaaa gataaaactg tagagtcctg
      961 ttgtcaaaat actcaattgt cctttagacc atgtctaact gttcatttat atgattctct
     1021 aaaacactga tattattgta gtactataga ttatattatt cgtagagtaa agtttaaata
     1081 tatgtataaa gatagataaa ctgcacttca aacaagtgtg acaaaaaaaa tatgtggtaa
     1141 ttttttataa cttagacatg caatgctcat tatctctaga gaggggcacg accgggtcac
     1201 gctgcactgc aggcatgcaa gctgatctga gcttggcact ggccgtcgtt ttacaacgtc
     1261 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1321 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1381 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1441 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     1501 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1561 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1621 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1681 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1801 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1921 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2041 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2101 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2221 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2401 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2521 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2701 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2761 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2821 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2881 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2941 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3001 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3061 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3121 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3181 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3241 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3301 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3361 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3421 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3481 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3541 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3601 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3661 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3721 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     3781 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     3841 gaaacagcta tgacatgatt acgaattcgc aaaacacacc tagactagat ttgttttgct
     3901 aacccaattg atattaatta tatatgatta atatttatat gtatatggat ttggttaatg
     3961 aaatgcatct ggttcatcaa agaattataa agacacgtga cattcattta ggataagaaa
     4021 tatggatgat ctctttctct tttattcaga taactagtaa ttacacataa cacacaactt
     4081 tgatgcccac attatagtga ttagcatgtc actatgtgtg catcctttta tttcatacat
     4141 taattaagtt ggccaatcca gaagatggac aagtctaggt taaccatggt ggatcctcta
     4201 gagtcaccgc ttccctatag ctttgcgaag agcctcgctg atcatccagg gcacgacggc
     4261 accggagcca gcaaactgag ctcgtatctt aagccccatg aagccatctg gcgctgaagc
     4321 ctgaaccgag aggacgtcca aatggaggct cttgatggcg tcgaacactc gcgtcatcag
     4381 gagctcctcc caccggcact gcacctccag gagcacgtcc ttgtccgaga cggtgacggt
     4441 gacgttgctg gtgccggcgt ccatggtgag gaccgggggg cgctccacgt cgtctctgcc
     4501 gagctctggg ctcttcctct tggagcccgc gcagacctcc ttcctcacac tctcattatt
     4561 gccacgggag ggccttgtta ttagccttgt cgtcgtttcg gatgggcgcg acgcaggttc
     4621 cctactggac tccagctctt gcacccttct ctgaagctcc ttgaggtaga ctatcgtttc
     4681 ggcgaggatc gacgctttgt tcaccctgtg aatggacgga agcagtgact tgaggacgag
     4741 gaacatctcg ttgagcttct ctcgtcgctt tcgctccgac atgacgtggt tcttggtgcc
     4801 agtgccactc atttcctgtg ctgctcccgt cgcgccgcca cagctctccc aagcaccgcc
     4861 gccggccacc actttcttca gcaatctctg cggctcctcg atggccggca ctactgctgc
     4921 gggcgccgcg tcgtcggagc acgacgactg ctgcgaggac ctcgtccaag ccatgaaact
     4981 cgtcgcgcga gagccgtaga cgggtgccct tgaggcgtcg gcggcgacgt tagcggtagc
     5041 cctgtccacc ggaggcggcg ctggctgcgg ggaagagcag gggacctcga aattggacgc
     5101 gtccacggtc cagccgtcct ctagcggtag tggtagcgcc tgcaggtcca tttcgtcgca
     5161 gaggctgtag aactcctcga tctccttggt gatgtgctcc aggcttgcgt cgtctgacag
     5221 ggcctcggct tctcttagtc ttagctcctc ctcctgcccg tgtcccccgg cggcggtcat
     5281 cgcctctatg tcgtccatgc cattattgtg gtcgagttcc tcgaacgcaa acgtgccgtc
     5341 gtctgctgcg gcctcgccgg tctcgtttgc tcgtcctgac gggctggagc tcgggcactg
     5401 cggctcccag aaagccgcgg ttgctcggct gaccaagtcc ggggcctccg gcaccgtgtc
     5461 agttgtacca agctcaagca cgccgcccat aaccgggatg cagaggattg actgaatgga
     5521 cgcgctcttg gccaggagcg cgcgggggaa ggctttgctg ccggcgaggt gcgcgttgca
     5581 cagccagaca tgctcgtcgc tcgcgaaact cctgccgggc aacccttggc ctggccggaa
     5641 ggcgtaggtc atggagacca cgtagtacca ctcggtgtcg ccgaggtcct ccggcgacag
     5701 agagccggcc ggccgcgcag gcgcagcgcg gcggtcgccc tcgcccgaca ggagggcctc
     5761 gtagagctcc cggagctggt cgctcctctg catgacgagc tggtcggatg tcagctccac
     5821 ggagttggag atcttccgcg tcttcacctc gccgttgtag aacccgtccg tccacgtcag
     5881 cacccttggt tgagtgtctg aaatggacca gaagagggcg tagctccagt tgatgctcct
     5941 ggcggctgca gcaagctggc tcctcatcag ctgcctctca gcaggtcgtt gcagcagttc
     6001 ttccgcctgc tgaactcggg aagctgaaag cgccatggtg gatcctctag agtcgacctg
     6061 cagaagtaac accaaacaac agggtgagca tcgacaaaag aaacagtacc aagcaaataa
     6121 atagcgtatg aaggcagggc taaaaaaatc cacatatagc tgctgcatat gccatcatcc
     6181 aagtatatca agatcaaaat aattataaaa catacttgtt tattataata gataggtact
     6241 caaggttaga gcatatgaat agatgctgca tatgccatca tgtatatgca tcagtaaaac
     6301 ccacatcaac atgtatacct atcctagatc gatatttcca tccatcttaa actcgtaact
     6361 atgaagatgt atgacacaca catacagttc caaaattaat aaatacacca ggtagtttga
     6421 aacagtattc tactccgatc tagaacgaat gaacgaccgc ccaaccacac cacatcatca
     6481 caaccaagcg aacaaaaagc atctctgtat atgcatcagt aaaacccgca tcaacatgta
     6541 tacctatcct agatcgatat ttccatccat catcttcaat tcgtaactat gaatatgtat
     6601 ggcacacaca tacagatcca aaattaataa atccaccagg tagtttgaaa cagaattcta
     6661 ctccgatcta gaacgaccgc ccaaccagac cacatcatca caaccaagac aaaaaaaagc
     6721 atgaaaagat gacccgacaa acaagtgcac ggcatatatt gaaataaagg aaaagggcaa
     6781 accaaaccct atgcaacgaa acaaaaaaaa tcatgaaatc gatcccgtct gcggaac
//
