LOCUS       sequence-244164-        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     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      3288..3541
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3590..3606)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3590..3606)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3603..3625)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3614..3630
                     /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(3638..3644,3645..3662,3663..3668))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3683..3704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3822..3839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3993..4581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4073..4092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4752..5567)
                     /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(4752..5567)
                     /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     5475..5494
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5568..5672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5740..5758
                     /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 aaaatgatta cgaccagaaa
     1861 ggtggctata tacgttcgtg tcagcactac taatcaggct gaggaagggt atagtattca
     1921 aggtcaaatc gattcactaa taaagtactg cgaagctatg ggctggatca tctatgagga
     1981 atatactgat gccgggttca gtggagggaa gattgatagg cccgctatga gtaaattaat
     2041 cacggatgca aagcacaaga gatttgatac gattcttgtc tataaacttg acagattaag
     2101 tagatccgtc agggacacgt tatacctggt gaaggatgtc ttcaatcaga acaatataca
     2161 cttcgtgagc ctacaggaaa acatcgatac ttcctcagcg atgggtaatc tattcttgac
     2221 cctcttatca gcgatagctg agtttgagcg agaacaaatt acggagcgaa tgacgatggg
     2281 caagattggg cgagccaaat ctggtaagac aatggcttgg acatatactc ctttcggata
     2341 cgactacaac aaggagaagg gagagctcat ccttgaccca gccaaagctc ctatcgtgaa
     2401 gatgatttac accgactatt taaaagggat gtccatacag aaaattgttg acaagctgaa
     2461 taagatggat tacaatggta aagattgcac ttggttcccg catggcgtga agcacctcct
     2521 agacaatcca gtctactatg gaatgacaag gtataacaac aaattattcc ctggaaatca
     2581 tcagcctatt atcaccaagg aactattcga caagacccag agagagagac agaggcgacg
     2641 attgggcatt gaagagaatc attacacaat tccattccag gctaagtaca tgctgtctaa
     2701 attcttgcga tgcaggcagt gcggttcaag gatggggctc gaacttggga gacctcgtaa
     2761 gaaggaaggt aaaaggtcaa agaaatacta ttgtctgaat tctagaccta agaggacggc
     2821 ttcctgcgac actcccctat acgatgctga gacactggaa gattacgtgc tgcatgaaat
     2881 tgccaaaatc caaaaggatc cgtcaatcgc gagtcgacaa aaacatatcg aggatcacga
     2941 gttaaagtac aagcgtgaac gtattgaagc caatataaat aaaaccgtaa atcaactgtc
     3001 caagttgaac aatttgtacc taaacgactt gatcaccttg gaggacctca agactcagac
     3061 gaacacactt atagcgaaga aacgtcttct cgaaaacgaa cttgataaga catgcgacaa
     3121 cgatgacgaa cttgataggc aggaaactat tgccgacttc ctggctctac ctgatgtgtg
     3181 gacgatggac tatgagggcc agaagtatgc tgttgagttg ttagtccaga gagttaaagt
     3241 ggacagggat aacatagata tacattggac cttttaagaa tttccccgat cgttcaaaca
     3301 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     3361 aatttctgtt gaattacgtt aagcatgtaa tatttaacat gtaatgcatg acgttattta
     3421 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     3481 aaatatcgcg cgcaaacttg gataaattat cgcgcgcggt gtcatctatg aggactagat
     3541 cggggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg tcatagctgt
     3601 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3661 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3721 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3781 cggggagagg cggtttgcgt attgggcgcg gccgccgctt cctcgctcac tgactcgctg
     3841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4741 tggtctgaca gttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     4801 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     4861 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     4921 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     4981 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     5041 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     5101 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     5161 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     5221 gaatcaggat attcttctaa tacctggaat gctgttttcc cagggatcgc agtggtgagt
     5281 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     5341 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     5401 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     5461 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     5521 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt
     5581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5701 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5761 tttcgtc
//
