LOCUS       sequence-250850-        5719 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  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 5719)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-AtINT9
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(71..93)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     193..212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     406..428
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     420..437
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     421..437
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3282..3535
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3584..3600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3584..3600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3597..3619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3608..3624
                     /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(3632..3638,3639..3656,3657..3662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3677..3698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3816..3833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4746..5561)
                     /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(4746..5561)
                     /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     5469..5488
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5562..5666)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
       61 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      121 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
      181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      241 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
      301 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      361 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      421 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gatatcggat
      481 cccaaaattt agaacgaact taattatgat ctcaaataca ttgatacata tctcatctag
      541 atctaggtta tcattatgta agaaagtttt gacgaatatg gcacgacaaa atggctagac
      601 tcgatgtaat tggtatctca actcaacatt atacttatac caaacattag ttagacaaaa
      661 tttaaacaac tattttttat gtatgcaaga gtcagcatat gtataattga ttcagaatcg
      721 ttttgacgag ttcggatgta gtagtagcca ttatttaatg tacatactaa tcgtgaatag
      781 tgaatatgat gaaacattgt atcttattgt ataaatatcc ataaacacat catgaaagac
      841 actttctttc acggtctgaa ttaattatga tacaattcta atagaaaacg aattaaatta
      901 cgttgaattg tatgaaatct aattgaacaa gccaaccacg acgacgacta acgttgcctg
      961 gattgactcg gtttaagtta accactaaaa aaacggagct gtcatgtaac acgcggatcg
     1021 agcaggtcac agtcatgaag ccatcaaagc aaaagaacta atccaagggc tgagatgatt
     1081 aattagttta aaaattagtt aacacgaggg aaaaggctgt ctgacagcca ggtcacgtta
     1141 tctttacctg tggtcgaaat gattcgtgtc tgtcgatttt aattattttt ttgaaaggcc
     1201 gaaaataaag ttgtaagaga taaacccgcc tatataaatt catatatttt cctctccgct
     1261 ttgaattgtc tcgttgtcct cctcactttc atcagccgtt ttgaatctcc ggcgacttga
     1321 cagagaagaa caaggaagaa gactaagaga gaaagtaaga gataatccag gagattcatt
     1381 ctccgttttg aatcttcctc aatctcatct tcttccgctc tttctttcca aggtaatagg
     1441 aactttctgg atctacttta tttgctggat ctcgatcttg ttttctcaat ttccttgaga
     1501 tctggaattc gtttaatttg gatctgtgaa cctccactaa atcttttggt tttactagaa
     1561 tcgatctaag ttgaccgatc agttagctcg attatagcta ccagaatttg gcttgacctt
     1621 gatggagaga tccatgttca tgttacctgg gaaatgattt gtatatgtga attgaaatct
     1681 gaactgttga agttagattg aatctgaaca ctgtcaatgt tagattgaat ctgaacactg
     1741 tttaaggtta gatgaagttt gtgtatagat tcttcgaaac tttaggattt gtagtgtcgt
     1801 acgttgaaca gaaagctatt tctgattcaa tcagggttta tttgactgta ttgaactctt
     1861 tttgtgtgtt tgcagctcat aaaaaatgaa ggtcgctata tacaccagag taagcactct
     1921 ggagcaaaag gagaaaggtc attctataga agaacaggag aggaagctca gagcatactc
     1981 cgatataaac gattggaaaa ttcataaagt gtatacagac gccggctatt ccggggcgaa
     2041 gaaagaccga ccagcactac aagagatgtt gaacgaaata gataacttcg atttggtgct
     2101 cgtttataag ctcgaccgac tcactcgttc tgtcaaagac ctacttgaaa ttctcgagct
     2161 atttgagaat aaaaatgtac tctttcgatc tgcgaccgaa gtctatgata ccacatcagc
     2221 catgggacga cttttcgtga cgttggtggg cgcgatggca gagtgggaaa gaacgactat
     2281 tcaagaaagg accgcaatgg gaagacgtgc cagtgccaga aagggtcttg cgaaaacggt
     2341 acctcctttc tattatgaca gggtcaatga caagttcgtg cctaatgagt acaagaaagt
     2401 gttgcgtttc gccgtggaag aagcgaagaa agggactagc cttagagaaa taacaatcaa
     2461 actgaataac agcaagtaca aggcgccact cggaaaaaat tggcacaggt ccgtaatcgg
     2521 aaacgcactc acctcacctg ttgctagagg ccatctagta tttggggaca ttttcgtgga
     2581 gaacactcac gaggcgataa tatcagaaga ggagtacgag gaaattaagc ttagaatctc
     2641 tgagaagacg aactcaacaa tcgttaaaca taatgcgatc ttcaggagta aactactatg
     2701 tccaaactgc aatcaaaagc ttaccctgaa taccgttaaa catacaccca agaataagga
     2761 agtgtggtat tcaaagctat atttctgcag taactgtaag aatacgaaaa ataagaatgc
     2821 ttgtaacatt gacgaaggtg aggttttaaa gcaattctat aattatctaa agcaatttga
     2881 tctgacttcc tataaaatcg agaaccagcc aaaggaaatc gaagatgtgg gtatcgacat
     2941 tgagaagttg aggaaagagc gagcgagatg ccagacgctc ttcattgaag gtatgatgga
     3001 caaagatgag gcttttccta ttatatcacg aatcgacaag gagatacacg agtatgagaa
     3061 aaggaaagat aacgacaaag ggaaaacatt caattatgaa aaaatcaaga actttaagta
     3121 cagtcttctc aatgggtggg aactcatgga ggatgaactt aagaccgagt tcataaagat
     3181 ggcaatcaaa aatatacatt ttgaatatgt gaaagggata aaggggaaga ggcaaaattc
     3241 tttgaagatc acgggcatag aattttatta agaatttccc cgatcgttca aacatttggc
     3301 aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc
     3361 tgttgaatta cgttaagcat gtaatattta acatgtaatg catgacgtta tttatgagat
     3421 gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat
     3481 cgcgcgcaaa cttggataaa ttatcgcgcg cggtgtcatc tatgaggact agatcggggc
     3541 ccgtcgactg cagaggcctg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     3601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3781 gaggcggttt gcgtattggg cgcggccgcc gcttcctcgc tcactgactc gctgcgctcg
     3841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4741 gacagttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     4801 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     4861 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     4921 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     4981 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     5041 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     5101 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     5161 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     5221 ggatattctt ctaatacctg gaatgctgtt ttcccaggga tcgcagtggt gagtaaccat
     5281 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     5341 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     5401 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     5461 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     5521 cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat
     5581 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5641 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5701 gaaaccatta ttatcatga
//
