LOCUS       sequence-378786-        5758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 13..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5758)
  AUTHORS   Gomide MS, Sales TT, Barros LRC, Limia CG, de Oliveira MA, 
            Florentino LH, Barros LMG, Robledo ML, Jose GPC, Almeida MSM, Lima 
            RN, Rehen SK, Lacorte C, Melo EO, Murad AM, Bonamino MH, Coelho CM, 
            Rech E
  TITLE     Genetic switches designed for eukaryotic cells and controlled by 
            serine integrases.
  JOURNAL   Commun Biol. 2020 May 22;3(1):255. doi: 10.1038/s42003-020-0971-8.
  PUBMED    32444777
REFERENCE   2  (bases 1 to 5758)
  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: "2020-05-22"; volume: "3(1):255. doi"; pages: " 
            10.1038/s42003-020-0971-8"
COMMENT     Sequence Label: INCTbiosyn-pAct-AtINT13
FEATURES             Location/Qualifiers
     source          1..5758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..41
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(79..101)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     201..220
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     414..436
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     428..445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     429..445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3329..3582
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3631..3647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3631..3647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3644..3666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3655..3671
                     /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(3679..3685,3686..3703,3704..3709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3724..3745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3863..3880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4034..4622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4793..5608)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4793..5608)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5516..5535
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5609..5713)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
       61 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      121 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      181 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
      241 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
      301 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
      361 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      421 acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa tgcatctaga
      481 tatcggatcc caaaatttag aacgaactta attatgatct caaatacatt gatacatatc
      541 tcatctagat ctaggttatc attatgtaag aaagttttga cgaatatggc acgacaaaat
      601 ggctagactc gatgtaattg gtatctcaac tcaacattat acttatacca aacattagtt
      661 agacaaaatt taaacaacta ttttttatgt atgcaagagt cagcatatgt ataattgatt
      721 cagaatcgtt ttgacgagtt cggatgtagt agtagccatt atttaatgta catactaatc
      781 gtgaatagtg aatatgatga aacattgtat cttattgtat aaatatccat aaacacatca
      841 tgaaagacac tttctttcac ggtctgaatt aattatgata caattctaat agaaaacgaa
      901 ttaaattacg ttgaattgta tgaaatctaa ttgaacaagc caaccacgac gacgactaac
      961 gttgcctgga ttgactcggt ttaagttaac cactaaaaaa acggagctgt catgtaacac
     1021 gcggatcgag caggtcacag tcatgaagcc atcaaagcaa aagaactaat ccaagggctg
     1081 agatgattaa ttagtttaaa aattagttaa cacgagggaa aaggctgtct gacagccagg
     1141 tcacgttatc tttacctgtg gtcgaaatga ttcgtgtctg tcgattttaa ttattttttt
     1201 gaaaggccga aaataaagtt gtaagagata aacccgccta tataaattca tatattttcc
     1261 tctccgcttt gaattgtctc gttgtcctcc tcactttcat cagccgtttt gaatctccgg
     1321 cgacttgaca gagaagaaca aggaagaaga ctaagagaga aagtaagaga taatccagga
     1381 gattcattct ccgttttgaa tcttcctcaa tctcatcttc ttccgctctt tctttccaag
     1441 gtaataggaa ctttctggat ctactttatt tgctggatct cgatcttgtt ttctcaattt
     1501 ccttgagatc tggaattcgt ttaatttgga tctgtgaacc tccactaaat cttttggttt
     1561 tactagaatc gatctaagtt gaccgatcag ttagctcgat tatagctacc agaatttggc
     1621 ttgaccttga tggagagatc catgttcatg ttacctggga aatgatttgt atatgtgaat
     1681 tgaaatctga actgttgaag ttagattgaa tctgaacact gtcaatgtta gattgaatct
     1741 gaacactgtt taaggttaga tgaagtttgt gtatagattc ttcgaaactt taggatttgt
     1801 agtgtcgtac gttgaacaga aagctatttc tgattcaatc agggtttatt tgactgtatt
     1861 gaactctttt tgtgtgtttg cagctcataa aaaatggctg tcggaattta catacgagtg
     1921 agtactcagg agcaggccag tgaggggcat tccatagaaa gccaaaagaa aaaattggcg
     1981 tcttactgcg agattcaggg gtgggatgac tataggtttt atattgagga gggaatatcc
     2041 gggaagaata caaatcgtcc aaagcttaaa ctccttatgg agcatattga aaagggtaaa
     2101 attaatatac tactcgtgta ccgattagac cgtctaacga ggtcagtaat cgatttgcac
     2161 aagttattga attttctgca ggaacacggt tgcgccttca agtccgcaac tgaaacttac
     2221 gacacaacca ccgcaaacgg aaggatgagt atggggattg tatcactcct tgcccagtgg
     2281 gagacggaaa atatgagcga acgtataaaa ctgaatctcg aacacaaagt actagtagag
     2341 ggggaaaggg ttggggctat cccctacgga tttgatctaa gtgatgacga aaagctagtg
     2401 aaaaatgaaa aaagtgcaat tctgctggat atggtggagc gagtagaaaa tggatggtcc
     2461 gtcaatcgaa tcgtgaatta cctgaatctt acgaacaatg accgaaattg gagtccgaat
     2521 ggcgtcctgc gtctacttcg aaacccggcg ctctacgggg cgacccgttg gaacgataag
     2581 atagctgaga acacacacga aggaattatc agtaaagaga ggttcaatag attgcagcaa
     2641 atcctagcag atcgttctat tcatcacagg cgtgatgtta agggtaccta catttttcag
     2701 ggcgtattaa gatgtccggt ttgtgatcag acattgagcg ttaacagatt catcaagaag
     2761 cgaaaggatg gcacggagta ctgtggcgtt ttgtatagat gccagccctg tattaaacaa
     2821 aataaataca atcttgcaat tggcgaggca cgatttctga aggctctcaa cgagtatatg
     2881 tccacggtgg aatttcaaac tgttgaggac gaggtcatcc ctaagaagag tgagcgagag
     2941 atgctggaaa gtcaacttca acagatagct agaaagagag aaaagtatca aaaagcgtgg
     3001 gcttctgatt tgatgtctga cgacgagttt gaaaagctta tggttgagac ccgagagact
     3061 tacgatgaat gcaagcagaa actcgagtct tgcgaggacc ctatcaaaat cgacgaaact
     3121 tacttaaagg agatagttta tatgttccat caaaccttca acgacctgga gtccgagaaa
     3181 cagaaggagt tcatctcaaa gtttattcgt acaatacgtt acaccgtgaa ggaacagcaa
     3241 ccaataaggc cagacaagag caagactggg aaaggcaagc agaaagtcat aattactgag
     3301 gtagaattct atcaataaga atttccccga tcgttcaaac atttggcaat aaagtttctt
     3361 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     3421 taagcatgta atatttaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     3481 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatcgc gcgcaaactt
     3541 ggataaatta tcgcgcgcgg tgtcatctat gaggactaga tcggggcccg tcgactgcag
     3601 aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3841 tattgggcgc ggccgccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4681 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4741 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttagaaaa
     4801 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     4861 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     4921 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     4981 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     5041 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     5101 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     5161 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     5221 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     5281 atacctggaa tgctgttttc ccagggatcg cagtggtgag taaccatgca tcatcaggag
     5341 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     5401 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     5461 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     5521 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     5581 aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt
     5641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattat
//
