LOCUS       sequence-276302-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cassette for testing phiC31 Excisionase activity using a fluorescent
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  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 5316)
  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: pExcisionase-AntiDsRed
FEATURES             Location/Qualifiers
     source          1..5316
                     /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             2283..2960
                     /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(2463..2483)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2875..2898
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        complement(3018..5007)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     primer_bind     complement(5127..5147)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5127..5145)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5166..5182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5166..5182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5179..5201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5190..5206
                     /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(5214..5220,5221..5238,5239..5244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5259..5280
                     /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 gtgtggagta cgcgcccggg gagcccaaag gttaccccag ttggggcact
     2281 acatggcctc ctccgagaac gtcatcaccg agttcatgcg cttcaaggtg cgcatggagg
     2341 gcaccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2401 gccacaacac cgtgaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     2461 tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc
     2521 ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2581 aggacggcgg cgtggcgacc gtgacccagg actcctccct gcaggacggc tgcttcatct
     2641 acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtgatg cagaagaaga
     2701 ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     2761 agacccacaa ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagtcca
     2821 tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac gccaagctgg
     2881 acatcacctc ccacaacgag gactacacca tcgtggagca gtacgagcgc accgagggcc
     2941 gccaccacct gttcctgtag cggtgcgggt gccagggcgt gcccttgagt tctctcagtt
     3001 gggggcgtag atttaaatgc agaagtaaca ccaaacaaca gggtgagcat cgacaaaaga
     3061 aacagtacca agcaaataaa tagcgtatga aggcagggct aaaaaaatcc acatatagct
     3121 gctgcatatg ccatcatcca agtatatcaa gatcaaaata attataaaac atacttgttt
     3181 attataatag ataggtactc aaggttagag catatgaata gatgctgcat atgccatcat
     3241 gtatatgcat cagtaaaacc cacatcaaca tgtataccta tcctagatcg atatttccat
     3301 ccatcttaaa ctcgtaacta tgaagatgta tgacacacac atacagttcc aaaattaata
     3361 aatacaccag gtagtttgaa acagtattct actccgatct agaacgaatg aacgaccgcc
     3421 caaccacacc acatcatcac aaccaagcga acaaaaagca tctctgtata tgcatcagta
     3481 aaacccgcat caacatgtat acctatccta gatcgatatt tccatccatc atcttcaatt
     3541 cgtaactatg aatatgtatg gcacacacat acagatccaa aattaataaa tccaccaggt
     3601 agtttgaaac agaattctac tccgatctag aacgaccgcc caaccagacc acatcatcac
     3661 aaccaagaca aaaaaaagca tgaaaagatg acccgacaaa caagtgcacg gcatatattg
     3721 aaataaagga aaagggcaaa ccaaacccta tgcaacgaaa caaaaaaaat catgaaatcg
     3781 atcccgtctg cggaacggct agagccatcc caggattccc caaagagaaa cactggcaag
     3841 ttagcaatca gaacgtgtct gacgtacagg tcgcatccgt gtacgaacgc tagcagcacg
     3901 gatctaacac aaacacggat ctaacacaaa catgaacaga agtagaacta ccgggcccta
     3961 accatggacc ggaacgccga tctagagaag gtagagangg ggggggggga ggacgagcgg
     4021 cgtaccttga agcggaggtg ccgacgggtg gatttggggg agatctggtt gtgtgtgtgt
     4081 gcgctccgaa caacacgagg ttggggaaag agggtgtgga gggggtgtct atttattacg
     4141 gcgggcgagg aagggaaagc gaaggagcgg tgggaaagga atcccccgta gctgccggtg
     4201 ccgtgagagg aggaggaggc cgcctgccgt gccggctcac gtctgccgct ccgccacgca
     4261 atttctggat gccgacagcg gagcaagtcc aacggtggag cggaactctc gagaggggtc
     4321 cagaggcagc gacagagatg ccgtgccgtc tgcttcgctt ggcccgacgc gacgctgctg
     4381 gttcgctggt tggtgtccgt tagactcgtc gacggcgttt aacaggctgg cattatctac
     4441 tcgaaacaag aaaaatgttt ccttagtttt tttaatttct taaagggtat ttgtttaatt
     4501 tttagtcact ttattttatt ctattttata tctaaattat taaataaaaa aactaaaata
     4561 gagttttagt tttcttaatt tagaggctaa aatagaataa aatagatgta ctaaaaaaat
     4621 tagtctataa aaaccattaa ccctaaaccc taaatggatg tactaataaa atggatgaag
     4681 tattatatag gtgaagctat ttgcaaaaaa aaaggagaac acatgcacac taaaaagata
     4741 aaactgtaga gtcctgttgt caaaatactc aattgtcctt tagaccatgt ctaactgttc
     4801 atttatatga ttctctaaaa cactgatatt attgtagtac tatagattat attattcgta
     4861 gagtaaagtt taaatatatg tataaagata gataaactgc acttcaaaca agtgtgacaa
     4921 aaaaaatatg tggtaatttt ttataactta gacatgcaat gctcattatc tctagagagg
     4981 ggcacgaccg ggtcacgctg cactgcatgc tgcagcccgg gcgacgtcga ctgtttaaac
     5041 gaggccttaa ttaagggccc aattggccac gtacgtctag atctggcgcg cctcgaggat
     5101 cgatccatgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     5161 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5221 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5281 attgcgttgc gctcactgcc cgctttccag tcggga
//
