LOCUS       sequence-244168-        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     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3279..3532
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3581..3597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3581..3597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3594..3616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3605..3621
                     /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(3629..3635,3636..3653,3654..3659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3674..3695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3813..3830)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3984..4572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4064..4083)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4743..5558)
                     /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(4743..5558)
                     /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     5466..5485
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5559..5663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5731..5749
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tatcggatcc caaaatttag aacgaactta attatgatct caaatacatt
      481 gatacatatc tcatctagat ctaggttatc attatgtaag aaagttttga cgaatatggc
      541 acgacaaaat ggctagactc gatgtaattg gtatctcaac tcaacattat acttatacca
      601 aacattagtt agacaaaatt taaacaacta ttttttatgt atgcaagagt cagcatatgt
      661 ataattgatt cagaatcgtt ttgacgagtt cggatgtagt agtagccatt atttaatgta
      721 catactaatc gtgaatagtg aatatgatga aacattgtat cttattgtat aaatatccat
      781 aaacacatca tgaaagacac tttctttcac ggtctgaatt aattatgata caattctaat
      841 agaaaacgaa ttaaattacg ttgaattgta tgaaatctaa ttgaacaagc caaccacgac
      901 gacgactaac gttgcctgga ttgactcggt ttaagttaac cactaaaaaa acggagctgt
      961 catgtaacac gcggatcgag caggtcacag tcatgaagcc atcaaagcaa aagaactaat
     1021 ccaagggctg agatgattaa ttagtttaaa aattagttaa cacgagggaa aaggctgtct
     1081 gacagccagg tcacgttatc tttacctgtg gtcgaaatga ttcgtgtctg tcgattttaa
     1141 ttattttttt gaaaggccga aaataaagtt gtaagagata aacccgccta tataaattca
     1201 tatattttcc tctccgcttt gaattgtctc gttgtcctcc tcactttcat cagccgtttt
     1261 gaatctccgg cgacttgaca gagaagaaca aggaagaaga ctaagagaga aagtaagaga
     1321 taatccagga gattcattct ccgttttgaa tcttcctcaa tctcatcttc ttccgctctt
     1381 tctttccaag gtaataggaa ctttctggat ctactttatt tgctggatct cgatcttgtt
     1441 ttctcaattt ccttgagatc tggaattcgt ttaatttgga tctgtgaacc tccactaaat
     1501 cttttggttt tactagaatc gatctaagtt gaccgatcag ttagctcgat tatagctacc
     1561 agaatttggc ttgaccttga tggagagatc catgttcatg ttacctggga aatgatttgt
     1621 atatgtgaat tgaaatctga actgttgaag ttagattgaa tctgaacact gtcaatgtta
     1681 gattgaatct gaacactgtt taaggttaga tgaagtttgt gtatagattc ttcgaaactt
     1741 taggatttgt agtgtcgtac gttgaacaga aagctatttc tgattcaatc agggtttatt
     1801 tgactgtatt gaactctttt tgtgtgtttg cagctcataa aaaatggctg tcggaattta
     1861 catacgagtg agtactcagg agcaggccag tgaggggcat tccatagaaa gccaaaagaa
     1921 aaaattggcg tcttactgcg agattcaggg gtgggatgac tataggtttt atattgagga
     1981 gggaatatcc gggaagaata caaatcgtcc aaagcttaaa ctccttatgg agcatattga
     2041 aaagggtaaa attaatatac tactcgtgta ccgattagac cgtctaacga ggtcagtaat
     2101 cgatttgcac aagttattga attttctgca ggaacacggt tgcgccttca agtccgcaac
     2161 tgaaacttac gacacaacca ccgcaaacgg aaggatgagt atggggattg tatcactcct
     2221 tgcccagtgg gagacggaaa atatgagcga acgtataaaa ctgaatctcg aacacaaagt
     2281 actagtagag ggggaaaggg ttggggctat cccctacgga tttgatctaa gtgatgacga
     2341 aaagctagtg aaaaatgaaa aaagtgcaat tctgctggat atggtggagc gagtagaaaa
     2401 tggatggtcc gtcaatcgaa tcgtgaatta cctgaatctt acgaacaatg accgaaattg
     2461 gagtccgaat ggcgtcctgc gtctacttcg aaacccggcg ctctacgggg cgacccgttg
     2521 gaacgataag atagctgaga acacacacga aggaattatc agtaaagaga ggttcaatag
     2581 attgcagcaa atcctagcag atcgttctat tcatcacagg cgtgatgtta agggtaccta
     2641 catttttcag ggcgtattaa gatgtccggt ttgtgatcag acattgagcg ttaacagatt
     2701 catcaagaag cgaaaggatg gcacggagta ctgtggcgtt ttgtatagat gccagccctg
     2761 tattaaacaa aataaataca atcttgcaat tggcgaggca cgatttctga aggctctcaa
     2821 cgagtatatg tccacggtgg aatttcaaac tgttgaggac gaggtcatcc ctaagaagag
     2881 tgagcgagag atgctggaaa gtcaacttca acagatagct agaaagagag aaaagtatca
     2941 aaaagcgtgg gcttctgatt tgatgtctga cgacgagttt gaaaagctta tggttgagac
     3001 ccgagagact tacgatgaat gcaagcagaa actcgagtct tgcgaggacc ctatcaaaat
     3061 cgacgaaact tacttaaagg agatagttta tatgttccat caaaccttca acgacctgga
     3121 gtccgagaaa cagaaggagt tcatctcaaa gtttattcgt acaatacgtt acaccgtgaa
     3181 ggaacagcaa ccaataaggc cagacaagag caagactggg aaaggcaagc agaaagtcat
     3241 aattactgag gtagaattct atcaataaga atttccccga tcgttcaaac atttggcaat
     3301 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     3361 tgaattacgt taagcatgta atatttaaca tgtaatgcat gacgttattt atgagatggg
     3421 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatcgc
     3481 gcgcaaactt ggataaatta tcgcgcgcgg tgtcatctat gaggactaga tcggggcccg
     3541 tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3601 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3661 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3721 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3781 gcggtttgcg tattgggcgc ggccgccgct tcctcgctca ctgactcgct gcgctcggtc
     3841 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4381 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4681 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4741 agttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     4801 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     4861 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     4921 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     4981 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     5041 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     5101 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     5161 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     5221 tattcttcta atacctggaa tgctgttttc ccagggatcg cagtggtgag taaccatgca
     5281 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     5341 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     5401 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     5461 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     5521 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     5581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5701 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
