LOCUS       sequence-244165-        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(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      3513..3766
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3815..3831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3815..3831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3828..3850)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3839..3855
                     /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(3863..3869,3870..3887,3888..3893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3908..3929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4047..4064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4218..4806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4298..4317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4977..5792)
                     /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(4977..5792)
                     /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     5700..5719
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5793..5897)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5965..5983
                     /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 aaaatgccag gcatgacgac
     1861 agagaccgga cccgacccgg cagggttaat agacctgttc tgtagaaaaa gcaaagctgt
     1921 caaatctagg gctaatgggg cgggtcaaag gagaaaacag gaaatttcca tagcagcgca
     1981 ggagacgctc gggaggaagg tagccgcgtt gttaggaatg caagtcaggc atgtatggaa
     2041 ggaggtagga tctgccagca gattcagaaa gggaaaggcg agggatgacc agtccaaagc
     2101 tctaaaggcc cttgagtctg gagaggtggg cgcactgtgg tgctatcgtc tagatagatg
     2161 ggatagaggg ggtgcaggtg ctatccttaa gataatcgaa ccggaggacg gtatgcctcg
     2221 tcgattgttg tttggctggg acgaagatac aggcaggccg gtgctggaca gtactaacaa
     2281 acgagaccgt ggtgaactaa tacgacgagc agaagaggct cgtgaggagg cggaaaagct
     2341 tagtgagagg gtaagagata ctaaagcgca tcaaagagaa aacggggaat gggtaaatgc
     2401 tagggcgccg tacggactca gggttgtact tgtcacagta tccgatgaag aaggcgatga
     2461 gtacgatgaa cgaaagcttg cagcagacga tgaagacgct ggaggcccgg atggtctaac
     2521 gaaagcggag gccgcacgac tcgtgttcac acttcccgtc actgatcgac tatcctacgc
     2581 gggaacggct catgctatga atacccgaga aataccctct ccgactgggg ggccatggat
     2641 tgccgtaacg gtcagggata tgatacagaa ccccgcctac gcgggctggc agacaaccgg
     2701 gcgacaggac ggtaaacaac gaagacttac cttttataat ggtgaaggaa agcgtgttag
     2761 cgtgatgcac ggaccaccac ttgttacgga tgaggaacag gaggcagcga aggcagcggt
     2821 taagggagag gatggggtcg gagttccctt agacggttca gaccatgaca cgaggcgaaa
     2881 gcatctactc tctggccgaa tgcgatgtcc tggctgcggg ggctcctgtt catacagtgg
     2941 caatggctat agatgttgga gatcttcagt caaaggcggt tgccccgccc ccacgtatgt
     3001 cgcgaggaag tccgtagaag agtatgttgc ttttaggtgg gctgctaagc tcgctgcctc
     3061 agagccggat gatccttttg ttatagcggt tgcagatagg tgggcggcgt taacccatcc
     3121 tcaggcgagc gaagatgaga aatatgccaa agctgccgtt cgagaggcag agaaaaatct
     3181 aggccgtttg ctgcgtgatc gacaaaatgg agtctacgac gggccggcgg agcaattttt
     3241 cgcacccgca tatcaagagg cattaagcac acttcaggca gccaaggatg ccgttagtga
     3301 aagctcagcg agtgctgctg tagacgtgag ctggatcgta gacagtagcg attacgagga
     3361 gctatggtta agagctacgc cgacaatgag gaatgccata atagacacgt gtatagatga
     3421 gatttgggtt gcgaagggtc agaggggaag gccgttcgat ggggacgagc gagtaaagat
     3481 taaatgggca gcgcgtacat aagaatttcc ccgatcgttc aaacatttgg caataaagtt
     3541 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
     3601 acgttaagca tgtaatattt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     3661 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tcgcgcgcaa
     3721 acttggataa attatcgcgc gcggtgtcat ctatgaggac tagatcgggg cccgtcgact
     3781 gcagaggcct gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3841 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3901 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3961 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4021 tgcgtattgg gcgcggccgc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4081 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4141 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4201 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4261 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4321 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4381 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4441 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4501 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4561 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4621 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4681 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4741 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4801 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4861 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4921 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag
     4981 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     5041 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     5101 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     5161 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     5221 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     5281 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     5341 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     5401 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     5461 tctaatacct ggaatgctgt tttcccaggg atcgcagtgg tgagtaacca tgcatcatca
     5521 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     5581 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     5641 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     5701 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     5761 gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc
     5821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5941 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
