LOCUS       sequence-244163-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  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 5863)
  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-AtINT2
FEATURES             Location/Qualifiers
     source          1..5863
                     /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      3384..3637
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3686..3702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3686..3702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3699..3721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3710..3726
                     /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(3734..3740,3741..3758,3759..3764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3779..3800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3918..3935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4089..4677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4169..4188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4848..5663)
                     /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(4848..5663)
                     /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     5571..5590
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5664..5768)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5836..5854
                     /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 aaaatgccaa tagcgccaga
     1861 atttctctca cttgcttatc caggtcaaga gttccccgcg tacctttacg gacgagcttc
     1921 tagggatccg aaacgtaagg gaagatctgt tcaatctcaa ttagacgaag ggagagctac
     1981 gtgtctcgat gcagggtggc caattgctgg tgagttcaaa gatgtcgata gatcggcctc
     2041 cgcatatgct agacgtacac gtgacgaatt tgaggaaatg atagcaggaa ttcaggccgg
     2101 cgaatgtaga attctcgtgg cctttgaagc atccagatat tatcgagatt tagaggcgta
     2161 tgtcaggctt agacgagtat gtcgtgaagc cggagttctg ttgtgttaca acggtcaggt
     2221 ttacgattta agcaaatccg ccgaccgaaa agcgaccgca caggacgccg ttaatgcaga
     2281 gggtgaggca gatgatatta gagaacgtaa tttgagaaca actagattga atgcaaagag
     2341 aggaggcgct catggccccg tgcctgatgg gtataaaagg agatacgacc ctgattcagg
     2401 tgatttagtt gatcaaatcc cacatccaga tagagctggt ctgataacag aaatattccg
     2461 tagggcagca gcggccgagc ctctcgctgc tatatgtcga gacttaaacg agagaggaga
     2521 aacaactcat agaggtaaag catggcaaag gcaccatctg cacgctatcc tgagaaatcc
     2581 agcgtacatc ggtcatagga gacatcttgg cgtggataca ggtaaaggaa tgtgggcacc
     2641 aatatgtgat gatgaagact tcgctgaaac attccaagct gtccaggaaa tactctcttt
     2701 accaggaagg caactatcgc ccggaccaga agcccaacac ctacagactg gcattgcttt
     2761 atgcggtgag catccagacg agccaccact acgttcggtc actgtaagag gtagaactaa
     2821 ttacaactgt tctacacgat atgatgttgc gatgagggag gatcgtatgg atgcttttgt
     2881 ggaagaatca gttattactt ggcttgcgag tgatgaagcg gtggctgctt tcgaagataa
     2941 tacagatgat gagagaacga gaaaagccag aattcgactt aaagttctag aagagcaatt
     3001 agaagccgct caaaaacagg ctagaacact ccgtcctgat gggatgggga tgctcttgtc
     3061 tatcgactct cttgctggtc tcgaggccga acttacgcca cagatagaca aggcaaggca
     3121 agagagcaga tccctgcatg tacccgcact tttgcgtgac cttctaggca agccacgagc
     3181 agacgttgat cgtgcatgga acgaagctct aaccctccct cagagacgta tgatacttag
     3241 gatggttgtt acgattagac tattcaaggc gggttctcgt ggtgtcaggg caatagagcc
     3301 tggtcgaatt acgctcagct acgttggaga acctggattt aagccagtgg gtggtaacag
     3361 agcaaaacag tgagaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
     3421 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     3481 atgtaatatt taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     3541 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atcgcgcgca aacttggata
     3601 aattatcgcg cgcggtgtca tctatgagga ctagatcggg gcccgtcgac tgcagaggcc
     3661 tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3721 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3781 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3841 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3901 ggcgcggccg ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3961 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4501 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca
     4861 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     4921 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     4981 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     5041 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     5101 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     5161 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     5221 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     5281 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     5341 tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     5401 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     5461 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     5521 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     5581 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     5641 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     5701 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5761 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5821 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
