LOCUS       sequence-250848-        5992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 5..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  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 5992)
  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-AtINT5
FEATURES             Location/Qualifiers
     source          1..5992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..42)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     142..161
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     355..377
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     369..386
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     370..386
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3504..3757
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3806..3822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3806..3822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3819..3841)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3830..3846
                     /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(3854..3860,3861..3878,3879..3884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3899..3920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4038..4055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4209..4797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4289..4308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4968..5783)
                     /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(4968..5783)
                     /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     5691..5710
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5784..5888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5956..5974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
       61 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      121 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
      181 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
      241 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      301 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      361 cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacctcgcga atgcatctag
      421 atatcggatc ccaaaattta gaacgaactt aattatgatc tcaaatacat tgatacatat
      481 ctcatctaga tctaggttat cattatgtaa gaaagttttg acgaatatgg cacgacaaaa
      541 tggctagact cgatgtaatt ggtatctcaa ctcaacatta tacttatacc aaacattagt
      601 tagacaaaat ttaaacaact attttttatg tatgcaagag tcagcatatg tataattgat
      661 tcagaatcgt tttgacgagt tcggatgtag tagtagccat tatttaatgt acatactaat
      721 cgtgaatagt gaatatgatg aaacattgta tcttattgta taaatatcca taaacacatc
      781 atgaaagaca ctttctttca cggtctgaat taattatgat acaattctaa tagaaaacga
      841 attaaattac gttgaattgt atgaaatcta attgaacaag ccaaccacga cgacgactaa
      901 cgttgcctgg attgactcgg tttaagttaa ccactaaaaa aacggagctg tcatgtaaca
      961 cgcggatcga gcaggtcaca gtcatgaagc catcaaagca aaagaactaa tccaagggct
     1021 gagatgatta attagtttaa aaattagtta acacgaggga aaaggctgtc tgacagccag
     1081 gtcacgttat ctttacctgt ggtcgaaatg attcgtgtct gtcgatttta attatttttt
     1141 tgaaaggccg aaaataaagt tgtaagagat aaacccgcct atataaattc atatattttc
     1201 ctctccgctt tgaattgtct cgttgtcctc ctcactttca tcagccgttt tgaatctccg
     1261 gcgacttgac agagaagaac aaggaagaag actaagagag aaagtaagag ataatccagg
     1321 agattcattc tccgttttga atcttcctca atctcatctt cttccgctct ttctttccaa
     1381 ggtaatagga actttctgga tctactttat ttgctggatc tcgatcttgt tttctcaatt
     1441 tccttgagat ctggaattcg tttaatttgg atctgtgaac ctccactaaa tcttttggtt
     1501 ttactagaat cgatctaagt tgaccgatca gttagctcga ttatagctac cagaatttgg
     1561 cttgaccttg atggagagat ccatgttcat gttacctggg aaatgatttg tatatgtgaa
     1621 ttgaaatctg aactgttgaa gttagattga atctgaacac tgtcaatgtt agattgaatc
     1681 tgaacactgt ttaaggttag atgaagtttg tgtatagatt cttcgaaact ttaggatttg
     1741 tagtgtcgta cgttgaacag aaagctattt ctgattcaat cagggtttat ttgactgtat
     1801 tgaactcttt ttgtgtgttt gcagctcata aaaaatgcca ggcatgacga cagagaccgg
     1861 acccgacccg gcagggttaa tagacctgtt ctgtagaaaa agcaaagctg tcaaatctag
     1921 ggctaatggg gcgggtcaaa ggagaaaaca ggaaatttcc atagcagcgc aggagacgct
     1981 cgggaggaag gtagccgcgt tgttaggaat gcaagtcagg catgtatgga aggaggtagg
     2041 atctgccagc agattcagaa agggaaaggc gagggatgac cagtccaaag ctctaaaggc
     2101 ccttgagtct ggagaggtgg gcgcactgtg gtgctatcgt ctagatagat gggatagagg
     2161 gggtgcaggt gctatcctta agataatcga accggaggac ggtatgcctc gtcgattgtt
     2221 gtttggctgg gacgaagata caggcaggcc ggtgctggac agtactaaca aacgagaccg
     2281 tggtgaacta atacgacgag cagaagaggc tcgtgaggag gcggaaaagc ttagtgagag
     2341 ggtaagagat actaaagcgc atcaaagaga aaacggggaa tgggtaaatg ctagggcgcc
     2401 gtacggactc agggttgtac ttgtcacagt atccgatgaa gaaggcgatg agtacgatga
     2461 acgaaagctt gcagcagacg atgaagacgc tggaggcccg gatggtctaa cgaaagcgga
     2521 ggccgcacga ctcgtgttca cacttcccgt cactgatcga ctatcctacg cgggaacggc
     2581 tcatgctatg aatacccgag aaataccctc tccgactggg gggccatgga ttgccgtaac
     2641 ggtcagggat atgatacaga accccgccta cgcgggctgg cagacaaccg ggcgacagga
     2701 cggtaaacaa cgaagactta ccttttataa tggtgaagga aagcgtgtta gcgtgatgca
     2761 cggaccacca cttgttacgg atgaggaaca ggaggcagcg aaggcagcgg ttaagggaga
     2821 ggatggggtc ggagttccct tagacggttc agaccatgac acgaggcgaa agcatctact
     2881 ctctggccga atgcgatgtc ctggctgcgg gggctcctgt tcatacagtg gcaatggcta
     2941 tagatgttgg agatcttcag tcaaaggcgg ttgccccgcc cccacgtatg tcgcgaggaa
     3001 gtccgtagaa gagtatgttg cttttaggtg ggctgctaag ctcgctgcct cagagccgga
     3061 tgatcctttt gttatagcgg ttgcagatag gtgggcggcg ttaacccatc ctcaggcgag
     3121 cgaagatgag aaatatgcca aagctgccgt tcgagaggca gagaaaaatc taggccgttt
     3181 gctgcgtgat cgacaaaatg gagtctacga cgggccggcg gagcaatttt tcgcacccgc
     3241 atatcaagag gcattaagca cacttcaggc agccaaggat gccgttagtg aaagctcagc
     3301 gagtgctgct gtagacgtga gctggatcgt agacagtagc gattacgagg agctatggtt
     3361 aagagctacg ccgacaatga ggaatgccat aatagacacg tgtatagatg agatttgggt
     3421 tgcgaagggt cagaggggaa ggccgttcga tggggacgag cgagtaaaga ttaaatgggc
     3481 agcgcgtaca taagaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
     3541 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     3601 atgtaatatt taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     3661 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atcgcgcgca aacttggata
     3721 aattatcgcg cgcggtgtca tctatgagga ctagatcggg gcccgtcgac tgcagaggcc
     3781 tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3841 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3901 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3961 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4021 ggcgcggccg ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca
     4981 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     5041 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     5101 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     5161 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     5221 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     5281 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     5341 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     5401 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     5461 tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     5521 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     5581 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     5641 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     5701 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     5761 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5881 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5941 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tt
//
