LOCUS       sequence-276305-        5285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cassette for testing Cre recombinase activity using a DsRed2 
            fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5285)
  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 5285)
  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: pCre-AntiDsRed
FEATURES             Location/Qualifiers
     source          1..5285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    37..58
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(73..78,79..96,97..103)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    111..127
                     /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     116..138
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     135..151
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     135..151
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     170..190
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        172..190
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        310..2300
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    complement(2308..2341)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2342..3019)
                     /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(2404..2427)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     2819..2839
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     protein_bind    3020..3053
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      3068..3320
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3343..3414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3415..3483,3484..4275)
                     /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(3633..3652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4446..5034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4935..4954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5188..5205
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
       61 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      121 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
      181 tcactaaagg gaacaaaagc tgggtaccat ggatcgatcc tcgaggcgcg ccagatctag
      241 acgtacgtgg ccaattgggc ccttaattaa ggcctcgttt aaacagtcga cgtcgcccgg
      301 gctgcagcat gcagtgcagc gtgacccggt cgtgcccctc tctagagata atgagcattg
      361 catgtctaag ttataaaaaa ttaccacata ttttttttgt cacacttgtt tgaagtgcag
      421 tttatctatc tttatacata tatttaaact ttactctacg aataatataa tctatagtac
      481 tacaataata tcagtgtttt agagaatcat ataaatgaac agttagacat ggtctaaagg
      541 acaattgagt attttgacaa caggactcta cagttttatc tttttagtgt gcatgtgttc
      601 tccttttttt ttgcaaatag cttcacctat ataatacttc atccatttta ttagtacatc
      661 catttagggt ttagggttaa tggtttttat agactaattt ttttagtaca tctattttat
      721 tctattttag cctctaaatt aagaaaacta aaactctatt ttagtttttt tatttaataa
      781 tttagatata aaatagaata aaataaagtg actaaaaatt aaacaaatac cctttaagaa
      841 attaaaaaaa ctaaggaaac atttttcttg tttcgagtag ataatgccag cctgttaaac
      901 gccgtcgacg agtctaacgg acaccaacca gcgaaccagc agcgtcgcgt cgggccaagc
      961 gaagcagacg gcacggcatc tctgtcgctg cctctggacc cctctcgaga gttccgctcc
     1021 accgttggac ttgctccgct gtcggcatcc agaaattgcg tggcggagcg gcagacgtga
     1081 gccggcacgg caggcggcct cctcctcctc tcacggcacc ggcagctacg ggggattcct
     1141 ttcccaccgc tccttcgctt tcccttcctc gcccgccgta ataaatagac accccctcca
     1201 caccctcttt ccccaacctc gtgttgttcg gagcgcacac acacacaacc agatctcccc
     1261 caaatccacc cgtcggcacc tccgcttcaa ggtacgccgc tcgtcctncc cccccccccc
     1321 tctctacctt ctctagatcg gcgttccggt ccatggttag ggcccggtag ttctacttct
     1381 gttcatgttt gtgttagatc cgtgtttgtg ttagatccgt gctgctagcg ttcgtacacg
     1441 gatgcgacct gtacgtcaga cacgttctga ttgctaactt gccagtgttt ctctttgggg
     1501 aatcctggga tggctctagc cgttccgcag acgggatcga tttcatgatt ttttttgttt
     1561 cgttgcatag ggtttggttt gcccttttcc tttatttcaa tatatgccgt gcacttgttt
     1621 gtcgggtcat cttttcatgc ttttttttgt cttggttgtg atgatgtggt ctggttgggc
     1681 ggtcgttcta gatcggagta gaattctgtt tcaaactacc tggtggattt attaattttg
     1741 gatctgtatg tgtgtgccat acatattcat agttacgaat tgaagatgat ggatggaaat
     1801 atcgatctag gataggtata catgttgatg cgggttttac tgatgcatat acagagatgc
     1861 tttttgttcg cttggttgtg atgatgtggt gtggttgggc ggtcgttcat tcgttctaga
     1921 tcggagtaga atactgtttc aaactacctg gtgtatttat taattttgga actgtatgtg
     1981 tgtgtcatac atcttcatag ttacgagttt aagatggatg gaaatatcga tctaggatag
     2041 gtatacatgt tgatgtgggt tttactgatg catatacatg atggcatatg cagcatctat
     2101 tcatatgctc taaccttgag tacctatcta ttataataaa caagtatgtt ttataattat
     2161 tttgatcttg atatacttgg atgatggcat atgcagcagc tatatgtgga tttttttagc
     2221 cctgccttca tacgctattt atttgcttgg tactgtttct tttgtcgatg ctcaccctgt
     2281 tgtttggtgt tacttctgca tttaaattac cgttcgtata gcatacatta tacgaagtta
     2341 tctacaggaa caggtggtgg cggccctcgg tgcgctcgta ctgctccacg atggtgtagt
     2401 cctcgttgtg ggaggtgatg tccagcttgg cgtccacgta gtagtagccg ggcagctgca
     2461 cgggcttctt ggccatgtag atggacttga actccaccag gtagtggccg ccgtccttca
     2521 gcttcagggc cttgtgggtc tcgcccttca gcacgccgtc gcgggggtac aggcgctcgg
     2581 tggaggcctc ccagcccatg gtcttcttct gcatcacggg gccgtcggag gggaagttca
     2641 cgccgatgaa cttcaccttg tagatgaagc agccgtcctg cagggaggag tcctgggtca
     2701 cggtcgccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     2761 aggacagctt cttgtagtcg gggatgtcgg cggggtgctt cacgtacacc ttggagccgt
     2821 actggaactg gggggacagg atgtcccagg cgaagggcag ggggccgccc ttggtcacct
     2881 tcagcttcac ggtgttgtgg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     2941 actcgtggcc gttcacggtg ccctccatgc gcaccttgaa gcgcatgaac tcggtgatga
     3001 cgttctcgga ggaggccatt accgttcgta taatgtatgc tatacgaagt tatactagtg
     3061 gatccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     3121 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     3181 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     3241 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     3301 gtcatctatg ttactagatc gggaattcga tatcaagcta tacattcaaa tatgtatccg
     3361 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3421 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3481 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3541 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3601 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3661 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3721 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3781 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3841 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3901 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3961 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4021 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4081 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4141 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4201 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4261 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4321 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4381 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5101 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5161 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5221 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5281 aggtt
//
