LOCUS       sequence-276296-        5332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cassette for testing R recombinase activity using a fluorescent 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5332)
  AUTHORS   Cody JP, Graham ND, Zhao C, Swyers NC, Birchler JA
  TITLE     Site-specific recombinase genome engineering toolkit in maize.
  JOURNAL   Plant Direct. 2020 Mar 9;4(3):e00209. doi: 10.1002/pld3.209. 
            eCollection 2020 Mar.
  PUBMED    32166212
REFERENCE   2  (bases 1 to 5332)
  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: "Plant 
            Direct."; date: "2020-03-9"; volume: "4(3):e00209. doi"; pages: " 
            10.1002/pld3.209. eCollection 2020 Mar"
COMMENT     Sequence Label: pR-AntiDsRed
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..98)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(252..840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(332..351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1011..1802,1803..1871))
                     /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     1634..1653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1872..1943)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1966..2218)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             2291..2968
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2471..2491)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2883..2906
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        complement(3034..5023)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     primer_bind     complement(5143..5163)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5143..5161)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5182..5198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5182..5198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5195..5217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5206..5222
                     /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)."
     promoter        complement(join(5230..5236,5237..5254,5255..5260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5275..5296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
       61 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      121 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      181 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      241 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      301 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      361 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      421 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      481 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      541 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      601 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      661 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      721 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      781 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      961 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1021 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1081 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1141 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1201 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1261 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1321 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1381 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1441 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1501 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1561 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1621 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1681 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1741 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1801 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1861 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1921 atgagcggat acatatttga atgtatagct tgatatcgaa ttcccgatct agtaacatag
     1981 atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt
     2041 attaaatgta taattgcggg actctaatca taaaaaccca tctcataaat aacgtcatgc
     2101 attacatgtt aattattaca tgcttaacgt aattcaacag aaattatatg ataatcatcg
     2161 caagaccggc aacaggattc aatcttaaga aactttattg ccaaatgttt gaacgatcgg
     2221 ggatccacta gtcgagatca tatcactgtg gacgttgatg aaagaatacg ttattctttc
     2281 atcaaatcgt atggcctcct ccgagaacgt catcaccgag ttcatgcgct tcaaggtgcg
     2341 catggagggc accgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2401 ctacgagggc cacaacaccg tgaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     2461 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc
     2521 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2581 gaacttcgag gacggcggcg tggcgaccgt gacccaggac tcctccctgc aggacggctg
     2641 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtgatgca
     2701 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     2761 gaagggcgag acccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     2821 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggacgc
     2881 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcac
     2941 cgagggccgc caccacctgt tcctgtagac gatttgatga aagaataacg tattctttca
     3001 tcaacgtcca cagtgatatg atctcgattt aaatgcagaa gtaacaccaa acaacagggt
     3061 gagcatcgac aaaagaaaca gtaccaagca aataaatagc gtatgaaggc agggctaaaa
     3121 aaatccacat atagctgctg catatgccat catccaagta tatcaagatc aaaataatta
     3181 taaaacatac ttgtttatta taatagatag gtactcaagg ttagagcata tgaatagatg
     3241 ctgcatatgc catcatgtat atgcatcagt aaaacccaca tcaacatgta tacctatcct
     3301 agatcgatat ttccatccat cttaaactcg taactatgaa gatgtatgac acacacatac
     3361 agttccaaaa ttaataaata caccaggtag tttgaaacag tattctactc cgatctagaa
     3421 cgaatgaacg accgcccaac cacaccacat catcacaacc aagcgaacaa aaagcatctc
     3481 tgtatatgca tcagtaaaac ccgcatcaac atgtatacct atcctagatc gatatttcca
     3541 tccatcatct tcaattcgta actatgaata tgtatggcac acacatacag atccaaaatt
     3601 aataaatcca ccaggtagtt tgaaacagaa ttctactccg atctagaacg accgcccaac
     3661 cagaccacat catcacaacc aagacaaaaa aaagcatgaa aagatgaccc gacaaacaag
     3721 tgcacggcat atattgaaat aaaggaaaag ggcaaaccaa accctatgca acgaaacaaa
     3781 aaaaatcatg aaatcgatcc cgtctgcgga acggctagag ccatcccagg attccccaaa
     3841 gagaaacact ggcaagttag caatcagaac gtgtctgacg tacaggtcgc atccgtgtac
     3901 gaacgctagc agcacggatc taacacaaac acggatctaa cacaaacatg aacagaagta
     3961 gaactaccgg gccctaacca tggaccggaa cgccgatcta gagaaggtag agaggggggg
     4021 gggggaggac gagcggcgta ccttgaagcg gaggtgccga cgggtggatt tgggggagat
     4081 ctggttgtgt gtgtgtgcgc tccgaacaac acgaggttgg ggaaagaggg tgtggagggg
     4141 gtgtctattt attacggcgg gcgaggaagg gaaagcgaag gagcggtggg aaaggaatcc
     4201 cccgtagctg ccggtgccgt gagaggagga ggaggccgcc tgccgtgccg gctcacgtct
     4261 gccgctccgc cacgcaattt ctggatgccg acagcggagc aagtccaacg gtggagcgga
     4321 actctcgaga ggggtccaga ggcagcgaca gagatgccgt gccgtctgct tcgcttggcc
     4381 cgacgcgacg ctgctggttc gctggttggt gtccgttaga ctcgtcgacg gcgtttaaca
     4441 ggctggcatt atctactcga aacaagaaaa atgtttcctt agttttttta atttcttaaa
     4501 gggtatttgt ttaattttta gtcactttat tttattctat tttatatcta aattattaaa
     4561 taaaaaaact aaaatagagt tttagttttc ttaatttaga ggctaaaata gaataaaata
     4621 gatgtactaa aaaaattagt ctataaaaac cattaaccct aaaccctaaa tggatgtact
     4681 aataaaatgg atgaagtatt atataggtga agctatttgc aaaaaaaaag gagaacacat
     4741 gcacactaaa aagataaaac tgtagagtcc tgttgtcaaa atactcaatt gtcctttaga
     4801 ccatgtctaa ctgttcattt atatgattct ctaaaacact gatattattg tagtactata
     4861 gattatatta ttcgtagagt aaagtttaaa tatatgtata aagatagata aactgcactt
     4921 caaacaagtg tgacaaaaaa aatatgtggt aattttttat aacttagaca tgcaatgctc
     4981 attatctcta gagaggggca cgaccgggtc acgctgcact gcatgctgca gcccgggcga
     5041 cgtcgactgt ttaaacgagg ccttaattaa gggcccaatt ggccacgtac gtctagatct
     5101 ggcgcgcctc gaggatcgat ccatggtacc cagcttttgt tccctttagt gagggttaat
     5161 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5221 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
     5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg ga
//
