LOCUS       sequence-276299-        5584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cassette for testing phiC31 Integrase activity using a DsRed2 
            fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  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 5584)
  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: pIntegrase-AntiDsRed
FEATURES             Location/Qualifiers
     source          1..5584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..22)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(176..764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(256..275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(935..1726,1727..1795))
                     /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     1558..1577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1796..1867)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1890..2142)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    2242..2341
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2371..3048
                     /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(2551..2571)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2963..2986
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(3050..3149)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    3169..3202
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        complement(3210..5199)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     primer_bind     complement(5319..5339)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5319..5337)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5358..5374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5358..5374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5371..5393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5382..5398
                     /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(5406..5412,5413..5430,5431..5436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5451..5472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
       61 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      121 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      361 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      601 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      661 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1861 ttgaatgtat agcttgatat cgaattcccg atctagtaac atagatgaca ccgcgcgcga
     1921 taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg
     1981 cgggactcta atcataaaaa cccatctcat aaataacgtc atgcattaca tgttaattat
     2041 tacatgctta acgtaattca acagaaatta tatgataatc atcgcaagac cggcaacagg
     2101 attcaatctt aagaaacttt attgccaaat gtttgaacga tcggggatcc actagtcccc
     2161 aactggggta acctttgagt tctctcagtt gggggttttg agacacgggc cagagctgca
     2221 gctagcatgg atctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
     2281 ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa aaaagcaggc
     2341 tcatttaact ttaagaagga gatatatacc atggcctcct ccgagaacgt catcaccgag
     2401 ttcatgcgct tcaaggtgcg catggagggc accgtgaacg gccacgagtt cgagatcgag
     2461 ggcgagggcg agggccgccc ctacgagggc cacaacaccg tgaagctgaa ggtgaccaag
     2521 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag
     2581 gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc
     2641 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggcgaccgt gacccaggac
     2701 tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt gaacttcccc
     2761 tccgacggcc ccgtgatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     2821 tacccccgcg acggcgtgct gaagggcgag acccacaagg ccctgaagct gaaggacggc
     2881 ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt gcagctgccc
     2941 ggctactact acgtggacgc caagctggac atcacctccc acaacgagga ctacaccatc
     3001 gtggagcagt acgagcgcac cgagggccgc caccacctgt tcctgtagta cccagctttc
     3061 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     3121 actatcagtc aaaataaaat cattatttgc catccagcta gcggtaatcg cgcccgggga
     3181 gcccaagggc acgccctggc acatttaaat gcagaagtaa caccaaacaa cagggtgagc
     3241 atcgacaaaa gaaacagtac caagcaaata aatagcgtat gaaggcaggg ctaaaaaaat
     3301 ccacatatag ctgctgcata tgccatcatc caagtatatc aagatcaaaa taattataaa
     3361 acatacttgt ttattataat agataggtac tcaaggttag agcatatgaa tagatgctgc
     3421 atatgccatc atgtatatgc atcagtaaaa cccacatcaa catgtatacc tatcctagat
     3481 cgatatttcc atccatctta aactcgtaac tatgaagatg tatgacacac acatacagtt
     3541 ccaaaattaa taaatacacc aggtagtttg aaacagtatt ctactccgat ctagaacgaa
     3601 tgaacgaccg cccaaccaca ccacatcatc acaaccaagc gaacaaaaag catctctgta
     3661 tatgcatcag taaaacccgc atcaacatgt atacctatcc tagatcgata tttccatcca
     3721 tcatcttcaa ttcgtaacta tgaatatgta tggcacacac atacagatcc aaaattaata
     3781 aatccaccag gtagtttgaa acagaattct actccgatct agaacgaccg cccaaccaga
     3841 ccacatcatc acaaccaaga caaaaaaaag catgaaaaga tgacccgaca aacaagtgca
     3901 cggcatatat tgaaataaag gaaaagggca aaccaaaccc tatgcaacga aacaaaaaaa
     3961 atcatgaaat cgatcccgtc tgcggaacgg ctagagccat cccaggattc cccaaagaga
     4021 aacactggca agttagcaat cagaacgtgt ctgacgtaca ggtcgcatcc gtgtacgaac
     4081 gctagcagca cggatctaac acaaacacgg atctaacaca aacatgaaca gaagtagaac
     4141 taccgggccc taaccatgga ccggaacgcc gatctagaga aggtagagag gggggggggg
     4201 gaggacgagc ggcgtacctt gaagcggagg tgccgacggg tggatttggg ggagatctgg
     4261 ttgtgtgtgt gtgcgctccg aacaacacga ggttggggaa agagggtgtg gagggggtgt
     4321 ctatttatta cggcgggcga ggaagggaaa gcgaaggagc ggtgggaaag gaatcccccg
     4381 tagctgccgg tgccgtgaga ggaggaggag gccgcctgcc gtgccggctc acgtctgccg
     4441 ctccgccacg caatttctgg atgccgacag cggagcaagt ccaacggtgg agcggaactc
     4501 tcgagagggg tccagaggca gcgacagaga tgccgtgccg tctgcttcgc ttggcccgac
     4561 gcgacgctgc tggttcgctg gttggtgtcc gttagactcg tcgacggcgt ttaacaggct
     4621 ggcattatct actcgaaaca agaaaaatgt ttccttagtt tttttaattt cttaaagggt
     4681 atttgtttaa tttttagtca ctttatttta ttctatttta tatctaaatt attaaataaa
     4741 aaaactaaaa tagagtttta gttttcttaa tttagaggct aaaatagaat aaaatagatg
     4801 tactaaaaaa attagtctat aaaaaccatt aaccctaaac cctaaatgga tgtactaata
     4861 aaatggatga agtattatat aggtgaagct atttgcaaaa aaaaaggaga acacatgcac
     4921 actaaaaaga taaaactgta gagtcctgtt gtcaaaatac tcaattgtcc tttagaccat
     4981 gtctaactgt tcatttatat gattctctaa aacactgata ttattgtagt actatagatt
     5041 atattattcg tagagtaaag tttaaatata tgtataaaga tagataaact gcacttcaaa
     5101 caagtgtgac aaaaaaaata tgtggtaatt ttttataact tagacatgca atgctcatta
     5161 tctctagaga ggggcacgac cgggtcacgc tgcactgcat gctgcagccc gggcgacgtc
     5221 gactgtttaa acgaggcctt aattaagggc ccaattggcc acgtacgtct agatctggcg
     5281 cgcctcgagg atcgatccat ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     5341 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5401 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5521 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5581 tccg
//
