LOCUS       sequence-250847-        5767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 4..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5767)
  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 5767)
  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-AtINT4
FEATURES             Location/Qualifiers
     source          1..5767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..109
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     303..325
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     317..334
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     318..334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3227..3480
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3529..3545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3529..3545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3542..3564)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3553..3569
                     /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(3577..3583,3584..3601,3602..3607))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3622..3643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3761..3778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3932..4520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4012..4031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4691..5506)
                     /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(4691..5506)
                     /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     5414..5433
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5507..5611)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5679..5697
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5735..5757)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
       61 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      121 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
      181 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      241 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
      301 ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta cctcgcgaat
      361 gcatctagat atcggatccc aaaatttaga acgaacttaa ttatgatctc aaatacattg
      421 atacatatct catctagatc taggttatca ttatgtaaga aagttttgac gaatatggca
      481 cgacaaaatg gctagactcg atgtaattgg tatctcaact caacattata cttataccaa
      541 acattagtta gacaaaattt aaacaactat tttttatgta tgcaagagtc agcatatgta
      601 taattgattc agaatcgttt tgacgagttc ggatgtagta gtagccatta tttaatgtac
      661 atactaatcg tgaatagtga atatgatgaa acattgtatc ttattgtata aatatccata
      721 aacacatcat gaaagacact ttctttcacg gtctgaatta attatgatac aattctaata
      781 gaaaacgaat taaattacgt tgaattgtat gaaatctaat tgaacaagcc aaccacgacg
      841 acgactaacg ttgcctggat tgactcggtt taagttaacc actaaaaaaa cggagctgtc
      901 atgtaacacg cggatcgagc aggtcacagt catgaagcca tcaaagcaaa agaactaatc
      961 caagggctga gatgattaat tagtttaaaa attagttaac acgagggaaa aggctgtctg
     1021 acagccaggt cacgttatct ttacctgtgg tcgaaatgat tcgtgtctgt cgattttaat
     1081 tatttttttg aaaggccgaa aataaagttg taagagataa acccgcctat ataaattcat
     1141 atattttcct ctccgctttg aattgtctcg ttgtcctcct cactttcatc agccgttttg
     1201 aatctccggc gacttgacag agaagaacaa ggaagaagac taagagagaa agtaagagat
     1261 aatccaggag attcattctc cgttttgaat cttcctcaat ctcatcttct tccgctcttt
     1321 ctttccaagg taataggaac tttctggatc tactttattt gctggatctc gatcttgttt
     1381 tctcaatttc cttgagatct ggaattcgtt taatttggat ctgtgaacct ccactaaatc
     1441 ttttggtttt actagaatcg atctaagttg accgatcagt tagctcgatt atagctacca
     1501 gaatttggct tgaccttgat ggagagatcc atgttcatgt tacctgggaa atgatttgta
     1561 tatgtgaatt gaaatctgaa ctgttgaagt tagattgaat ctgaacactg tcaatgttag
     1621 attgaatctg aacactgttt aaggttagat gaagtttgtg tatagattct tcgaaacttt
     1681 aggatttgta gtgtcgtacg ttgaacagaa agctatttct gattcaatca gggtttattt
     1741 gactgtattg aactcttttt gtgtgtttgc agctcataaa aaatgattac gaccagaaag
     1801 gtggctatat acgttcgtgt cagcactact aatcaggctg aggaagggta tagtattcaa
     1861 ggtcaaatcg attcactaat aaagtactgc gaagctatgg gctggatcat ctatgaggaa
     1921 tatactgatg ccgggttcag tggagggaag attgataggc ccgctatgag taaattaatc
     1981 acggatgcaa agcacaagag atttgatacg attcttgtct ataaacttga cagattaagt
     2041 agatccgtca gggacacgtt atacctggtg aaggatgtct tcaatcagaa caatatacac
     2101 ttcgtgagcc tacaggaaaa catcgatact tcctcagcga tgggtaatct attcttgacc
     2161 ctcttatcag cgatagctga gtttgagcga gaacaaatta cggagcgaat gacgatgggc
     2221 aagattgggc gagccaaatc tggtaagaca atggcttgga catatactcc tttcggatac
     2281 gactacaaca aggagaaggg agagctcatc cttgacccag ccaaagctcc tatcgtgaag
     2341 atgatttaca ccgactattt aaaagggatg tccatacaga aaattgttga caagctgaat
     2401 aagatggatt acaatggtaa agattgcact tggttcccgc atggcgtgaa gcacctccta
     2461 gacaatccag tctactatgg aatgacaagg tataacaaca aattattccc tggaaatcat
     2521 cagcctatta tcaccaagga actattcgac aagacccaga gagagagaca gaggcgacga
     2581 ttgggcattg aagagaatca ttacacaatt ccattccagg ctaagtacat gctgtctaaa
     2641 ttcttgcgat gcaggcagtg cggttcaagg atggggctcg aacttgggag acctcgtaag
     2701 aaggaaggta aaaggtcaaa gaaatactat tgtctgaatt ctagacctaa gaggacggct
     2761 tcctgcgaca ctcccctata cgatgctgag acactggaag attacgtgct gcatgaaatt
     2821 gccaaaatcc aaaaggatcc gtcaatcgcg agtcgacaaa aacatatcga ggatcacgag
     2881 ttaaagtaca agcgtgaacg tattgaagcc aatataaata aaaccgtaaa tcaactgtcc
     2941 aagttgaaca atttgtacct aaacgacttg atcaccttgg aggacctcaa gactcagacg
     3001 aacacactta tagcgaagaa acgtcttctc gaaaacgaac ttgataagac atgcgacaac
     3061 gatgacgaac ttgataggca ggaaactatt gccgacttcc tggctctacc tgatgtgtgg
     3121 acgatggact atgagggcca gaagtatgct gttgagttgt tagtccagag agttaaagtg
     3181 gacagggata acatagatat acattggacc ttttaagaat ttccccgatc gttcaaacat
     3241 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
     3301 atttctgttg aattacgtta agcatgtaat atttaacatg taatgcatga cgttatttat
     3361 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa
     3421 aatatcgcgc gcaaacttgg ataaattatc gcgcgcggtg tcatctatga ggactagatc
     3481 ggggcccgtc gactgcagag gcctgcatgc aagcttggcg taatcatggt catagctgtt
     3541 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3601 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3661 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3721 ggggagaggc ggtttgcgta ttgggcgcgg ccgccgcttc ctcgctcact gactcgctgc
     3781 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3841 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3901 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3961 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4021 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4081 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4141 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4201 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4261 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4321 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4381 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4441 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4501 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4561 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4621 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4681 ggtctgacag ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     4741 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     4801 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     4861 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     4921 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     4981 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     5041 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     5101 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     5161 aatcaggata ttcttctaat acctggaatg ctgttttccc agggatcgca gtggtgagta
     5221 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     5281 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     5341 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     5401 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     5461 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc
     5521 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5581 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5641 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5701 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5761 cggtcac
//
