LOCUS       sequence-250849-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase 7..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5877)
  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 5877)
  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-AtINT7
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2059..2312
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2327..2347)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2327..2345)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      2370..2427
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     CDS             complement(2585..3400)
                     /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(2585..3400)
                     /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     3308..3327
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3392..3411
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(3995..4540)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4075..4094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5008..5026
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5030..5073)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5079..5096
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5080..5096
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5103..5122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5103..5121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgcctatata aattcatata ttttcctctc cgctttgaat tgtctcgttg tcctcctcac
       61 tttcatcagc cgttttgaat ctccggcgac ttgacagaga agaacaagga agaagactaa
      121 gagagaaagt aagagataat ccaggagatt cattctccgt tttgaatctt cctcaatctc
      181 atcttcttcc gctctttctt tccaaggtaa taggaacttt ctggatctac tttatttgct
      241 ggatctcgat cttgttttct caatttcctt gagatctgga attcgtttaa tttggatctg
      301 tgaacctcca ctaaatcttt tggttttact agaatcgatc taagttgacc gatcagttag
      361 ctcgattata gctaccagaa tttggcttga ccttgatgga gagatccatg ttcatgttac
      421 ctgggaaatg atttgtatat gtgaattgaa atctgaactg ttgaagttag attgaatctg
      481 aacactgtca atgttagatt gaatctgaac actgtttaag gttagatgaa gtttgtgtat
      541 agattcttcg aaactttagg atttgtagtg tcgtacgttg aacagaaagc tatttctgat
      601 tcaatcaggg tttatttgac tgtattgaac tctttttgtg tgtttgcagc tcataaaaaa
      661 tgaaggtagc gatctatgtc cgtgtgtcaa ctgatgagca ggccaaggaa ggattttcca
      721 tcccagcgca gagagaaaga ttaagggcat tttgtgcctc acaaggctgg gagatcgtac
      781 aggagtatat cgaagagggg tggtctgcga aggatttgga cagacctcag atgcagagac
      841 tgcttaagga tataaaaaaa ggtaacatcg acattgtcct cgtatacagg ttggataggc
      901 taactaggtc cgttctggat ctttatctgc tcctgcaaac atttgagaag tacaacgtgg
      961 cttttcgtag tgctacggag gtatacgata cgtcaactgc aatgggccgt ttattcatta
     1021 ccttagtggc cgctctggca caatgggagc gtgaaaacct ggcagagagg gtaaaatttg
     1081 gaattgaaca gatgattgac gagggcaaga agcccggagg tcacagtccc tacggttata
     1141 aatttgataa ggacttcaac tgtactataa tcgaggagga agctgatgtc gtgcgtatga
     1201 tttacaggat gtactgcgat ggatatggct accgtagtat cgccgaccga cttaatgagt
     1261 tgatggtcaa acccaggata gccaaggaat ggaatcataa ttccgttcga gatattctta
     1321 caaacgatat ttatataggt acatacaggt ggggtgataa agtagtcccc aataatcatc
     1381 cgccaataat ctccgaaact ttgttcaaaa aagcacagaa ggaaaaagaa aagcgtggcg
     1441 ttgaccgaaa gcgagtcgga aagtttcttt ttacaggact gttgcaatgc gggaattgcg
     1501 ggggacacaa gatgcaaggg cacttcgata agcgtgagca aaagacttat taccgttgta
     1561 ctaaatgcca ccgtatcaca aacgagaaga acatactaga gccacttctt gacgagatac
     1621 agttgttaat tacgagtaaa gagtacttta tgagtaaatt tagcgacaga tacgatcaac
     1681 aggaggttgt agacgtgagc gcgctgacta aagaactgga gaagataaag aggcagaagg
     1741 agaagtggta cgatttgtat atggacgatc gaaatccgat ccccaaggaa gagctatttg
     1801 ccaaaataaa cgaattgaac aagaaagaag aggagatata tagtaagcta agtgaagtgg
     1861 aagaagataa agaaccggta gaagagaagt acaaccgtct ctccaaaatg attgatttta
     1921 aacaacagtt cgaacaagca aatgatttca caaaaaagga acttttgttc tctatcttcg
     1981 aaaagattgt aatctataga gagaaaggga aattaaagaa gatcacacta gattatactt
     2041 tgaagtaaga atttccccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     2101 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     2161 atatttaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     2221 caattataca tttaatacgc gatagaaaac aaaatatcgc gcgcaaactt ggataaatta
     2281 tcgcgcgcgg tgtcatctat gaggactaga tcgatcctcg aggttccctt tagtgagggt
     2341 taattgcgag cttggcgtaa tcatggtcat ccggcaaaaa agggcaaggt gtcaccaccc
     2401 tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact
     2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2521 atctcgagtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     2581 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     2641 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     2701 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     2761 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     2821 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     2881 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     2941 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3001 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3061 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3121 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3181 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3241 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3301 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3361 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3421 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     3481 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     3541 aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca
     3601 aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct
     3661 ggctggatga tggggcgatt caggcctggt atgagtcagc aacaccttct tcacgaggca
     3721 gacctcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     3781 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     3841 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     3901 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     3961 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     4021 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     4081 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     4141 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     4201 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     4261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     4321 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     4381 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     4441 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     4501 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg
     4561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4621 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4681 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4741 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4801 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4861 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4921 tgcaacttta tccgcctcca tccagtctat tccatggtgc cacctgacgt ctaagaaacc
     4981 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa
     5041 aaaaaatcct tagctttcgc taaggatgat ttctgcgttg taaaacgacg gccagtgaat
     5101 tgtaatacga ctcactatag ggcgacccgg ggataaaatt tagaacgaac ttaattatga
     5161 tctcaaatac attgatacat atctcatcta gatctaggtt atcattatgt aagaaagttt
     5221 tgacgaatat ggcacgacaa aatggctaga ctcgatgtaa ttggtatctc aactcaacat
     5281 tatacttata ccaaacatta gttagacaaa atttaaacaa ctatttttta tgtatgcaag
     5341 agtcagcata tgtataattg attcagaatc gttttgacga gttcggatgt agtagtagcc
     5401 attatttaat gtacatacta atcgtgaata gtgaatatga tgaaacattg tatcttattg
     5461 tataaatatc cataaacaca tcatgaaaga cactttcttt cacggtctga attaattatg
     5521 atacaattct aatagaaaac gaattaaatt acgttgaatt gtatgaaatc taattgaaca
     5581 agccaaccac gacgacgact aacgttgcct ggattgactc ggtttaagtt aaccactaaa
     5641 aaaacggagc tgtcatgtaa cacgcggatc gagcaggtca cagtcatgaa gccatcaaag
     5701 caaaagaact aatccaaggg ctgagatgat taattagttt aaaaattagt taacacgagg
     5761 gaaaaggctg tctgacagcc aggtcacgtt atctttacct gtggtcgaaa tgattcgtgt
     5821 ctgtcgattt taattatttt tttgaaaggc cgaaaataaa gttgtaagag ataaacc
//
