LOCUS       sequence-250845-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  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 5863)
  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-AtINT2
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..46)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     146..165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     359..381
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     373..390
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     374..390
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3379..3632
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3681..3697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3681..3697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3694..3716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3705..3721
                     /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(3729..3735,3736..3753,3754..3759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3774..3795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3913..3930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4084..4672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4164..4183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4843..5658)
                     /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(4843..5658)
                     /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     5566..5585
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5659..5763)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5831..5849
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
       61 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      121 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
      181 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
      241 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      301 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      361 cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacctc gcgaatgcat
      421 ctagatatcg gatcccaaaa tttagaacga acttaattat gatctcaaat acattgatac
      481 atatctcatc tagatctagg ttatcattat gtaagaaagt tttgacgaat atggcacgac
      541 aaaatggcta gactcgatgt aattggtatc tcaactcaac attatactta taccaaacat
      601 tagttagaca aaatttaaac aactattttt tatgtatgca agagtcagca tatgtataat
      661 tgattcagaa tcgttttgac gagttcggat gtagtagtag ccattattta atgtacatac
      721 taatcgtgaa tagtgaatat gatgaaacat tgtatcttat tgtataaata tccataaaca
      781 catcatgaaa gacactttct ttcacggtct gaattaatta tgatacaatt ctaatagaaa
      841 acgaattaaa ttacgttgaa ttgtatgaaa tctaattgaa caagccaacc acgacgacga
      901 ctaacgttgc ctggattgac tcggtttaag ttaaccacta aaaaaacgga gctgtcatgt
      961 aacacgcgga tcgagcaggt cacagtcatg aagccatcaa agcaaaagaa ctaatccaag
     1021 ggctgagatg attaattagt ttaaaaatta gttaacacga gggaaaaggc tgtctgacag
     1081 ccaggtcacg ttatctttac ctgtggtcga aatgattcgt gtctgtcgat tttaattatt
     1141 tttttgaaag gccgaaaata aagttgtaag agataaaccc gcctatataa attcatatat
     1201 tttcctctcc gctttgaatt gtctcgttgt cctcctcact ttcatcagcc gttttgaatc
     1261 tccggcgact tgacagagaa gaacaaggaa gaagactaag agagaaagta agagataatc
     1321 caggagattc attctccgtt ttgaatcttc ctcaatctca tcttcttccg ctctttcttt
     1381 ccaaggtaat aggaactttc tggatctact ttatttgctg gatctcgatc ttgttttctc
     1441 aatttccttg agatctggaa ttcgtttaat ttggatctgt gaacctccac taaatctttt
     1501 ggttttacta gaatcgatct aagttgaccg atcagttagc tcgattatag ctaccagaat
     1561 ttggcttgac cttgatggag agatccatgt tcatgttacc tgggaaatga tttgtatatg
     1621 tgaattgaaa tctgaactgt tgaagttaga ttgaatctga acactgtcaa tgttagattg
     1681 aatctgaaca ctgtttaagg ttagatgaag tttgtgtata gattcttcga aactttagga
     1741 tttgtagtgt cgtacgttga acagaaagct atttctgatt caatcagggt ttatttgact
     1801 gtattgaact ctttttgtgt gtttgcagct cataaaaaat gccaatagcg ccagaatttc
     1861 tctcacttgc ttatccaggt caagagttcc ccgcgtacct ttacggacga gcttctaggg
     1921 atccgaaacg taagggaaga tctgttcaat ctcaattaga cgaagggaga gctacgtgtc
     1981 tcgatgcagg gtggccaatt gctggtgagt tcaaagatgt cgatagatcg gcctccgcat
     2041 atgctagacg tacacgtgac gaatttgagg aaatgatagc aggaattcag gccggcgaat
     2101 gtagaattct cgtggccttt gaagcatcca gatattatcg agatttagag gcgtatgtca
     2161 ggcttagacg agtatgtcgt gaagccggag ttctgttgtg ttacaacggt caggtttacg
     2221 atttaagcaa atccgccgac cgaaaagcga ccgcacagga cgccgttaat gcagagggtg
     2281 aggcagatga tattagagaa cgtaatttga gaacaactag attgaatgca aagagaggag
     2341 gcgctcatgg ccccgtgcct gatgggtata aaaggagata cgaccctgat tcaggtgatt
     2401 tagttgatca aatcccacat ccagatagag ctggtctgat aacagaaata ttccgtaggg
     2461 cagcagcggc cgagcctctc gctgctatat gtcgagactt aaacgagaga ggagaaacaa
     2521 ctcatagagg taaagcatgg caaaggcacc atctgcacgc tatcctgaga aatccagcgt
     2581 acatcggtca taggagacat cttggcgtgg atacaggtaa aggaatgtgg gcaccaatat
     2641 gtgatgatga agacttcgct gaaacattcc aagctgtcca ggaaatactc tctttaccag
     2701 gaaggcaact atcgcccgga ccagaagccc aacacctaca gactggcatt gctttatgcg
     2761 gtgagcatcc agacgagcca ccactacgtt cggtcactgt aagaggtaga actaattaca
     2821 actgttctac acgatatgat gttgcgatga gggaggatcg tatggatgct tttgtggaag
     2881 aatcagttat tacttggctt gcgagtgatg aagcggtggc tgctttcgaa gataatacag
     2941 atgatgagag aacgagaaaa gccagaattc gacttaaagt tctagaagag caattagaag
     3001 ccgctcaaaa acaggctaga acactccgtc ctgatgggat ggggatgctc ttgtctatcg
     3061 actctcttgc tggtctcgag gccgaactta cgccacagat agacaaggca aggcaagaga
     3121 gcagatccct gcatgtaccc gcacttttgc gtgaccttct aggcaagcca cgagcagacg
     3181 ttgatcgtgc atggaacgaa gctctaaccc tccctcagag acgtatgata cttaggatgg
     3241 ttgttacgat tagactattc aaggcgggtt ctcgtggtgt cagggcaata gagcctggtc
     3301 gaattacgct cagctacgtt ggagaacctg gatttaagcc agtgggtggt aacagagcaa
     3361 aacagtgaga atttccccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     3421 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     3481 atatttaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     3541 caattataca tttaatacgc gatagaaaac aaaatatcgc gcgcaaactt ggataaatta
     3601 tcgcgcgcgg tgtcatctat gaggactaga tcggggcccg tcgactgcag aggcctgcat
     3661 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3721 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3781 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3841 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3901 ggccgccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4501 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttagaaaa actcatcgag
     4861 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     4921 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     4981 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     5041 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     5101 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     5161 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     5221 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     5281 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     5341 tgctgttttc ccagggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     5401 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     5461 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     5521 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     5581 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     5641 ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5701 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5761 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5821 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcg
//
