LOCUS       sequence-250853-        6363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes A. thaliana codon optimized Integrase phiC31..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6363)
  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 6363)
  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-AtINTphiC31
FEATURES             Location/Qualifiers
     source          1..6363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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)."
     primer_bind     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        228..246
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1674..3491
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     regulatory      2896..2905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      3516..3769
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3778..3797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3779..3797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3804..3821)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3804..3820)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3813..3835)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3962..4417)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4099..4120)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4311..4330
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4444..4548
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4549..4617,4618..5409)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4767..4786)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5580..6168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6069..6088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6322..6339
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac
      241 taaagggaac aaaagctgaa aatttagaac gaacttaatt atgatctcaa atacattgat
      301 acatatctca tctagatcta ggttatcatt atgtaagaaa gttttgacga atatggcacg
      361 acaaaatggc tagactcgat gtaattggta tctcaactca acattatact tataccaaac
      421 attagttaga caaaatttaa acaactattt tttatgtatg caagagtcag catatgtata
      481 attgattcag aatcgttttg acgagttcgg atgtagtagt agccattatt taatgtacat
      541 actaatcgtg aatagtgaat atgatgaaac attgtatctt attgtataaa tatccataaa
      601 cacatcatga aagacacttt ctttcacggt ctgaattaat tatgatacaa ttctaataga
      661 aaacgaatta aattacgttg aattgtatga aatctaattg aacaagccaa ccacgacgac
      721 gactaacgtt gcctggattg actcggttta agttaaccac taaaaaaacg gagctgtcat
      781 gtaacacgcg gatcgagcag gtcacagtca tgaagccatc aaagcaaaag aactaatcca
      841 agggctgaga tgattaatta gtttaaaaat tagttaacac gagggaaaag gctgtctgac
      901 agccaggtca cgttatcttt acctgtggtc gaaatgattc gtgtctgtcg attttaatta
      961 tttttttgaa aggccgaaaa taaagttgta agagataaac ccgcctatat aaattcatat
     1021 attttcctct ccgctttgaa ttgtctcgtt gtcctcctca ctttcatcag ccgttttgaa
     1081 tctccggcga cttgacagag aagaacaagg aagaagacta agagagaaag taagagataa
     1141 tccaggagat tcattctccg ttttgaatct tcctcaatct catcttcttc cgctctttct
     1201 ttccaaggta ataggaactt tctggatcta ctttatttgc tggatctcga tcttgttttc
     1261 tcaatttcct tgagatctgg aattcgttta atttggatct gtgaacctcc actaaatctt
     1321 ttggttttac tagaatcgat ctaagttgac cgatcagtta gctcgattat agctaccaga
     1381 atttggcttg accttgatgg agagatccat gttcatgtta cctgggaaat gatttgtata
     1441 tgtgaattga aatctgaact gttgaagtta gattgaatct gaacactgtc aatgttagat
     1501 tgaatctgaa cactgtttaa ggttagatga agtttgtgta tagattcttc gaaactttag
     1561 gatttgtagt gtcgtacgtt gaacagaaag ctatttctga ttcaatcagg gtttatttga
     1621 ctgtattgaa ctctttttgt gtgtttgcag ctcataaaaa caagcttcct aggatggaca
     1681 catacgcggg tgcctatgac aggcagtccc gtgaacgtga aaatagtagt gcggcttccc
     1741 ccgccacgca gcgttccgcc aatgaggaca aagcagcaga cctccaacga gaggttgagc
     1801 gagatggggg caggttcagg ttcgtaggcc acttctctga agcgcccggc acttccgcgt
     1861 tcggcaccgc ggagagaccc gagttcgagc gtatactaaa tgagtgccga gcgggtcgac
     1921 tcaacatgat aattgtatat gacgttagta ggttctccag actgaaagta atggatgcaa
     1981 taccaatagt atcagagctt ctagccttgg gcgttacgat agtgagcacc caggagggcg
     2041 tattccgaca agggaacgtt atggatctta tacatctaat tatgcgactt gatgccagtc
     2101 acaaggaatc ctccctcaag agcgccaaga ttcttgatac taagaaccta cagagggagc
     2161 tcgggggtta cgtcggggga aaagccccgt acggttttga acttgtcagt gaaaccaagg
     2221 agattacgag gaacgggagg atggtcaatg ttgtcataaa caagttggcg cactcaacta
     2281 ctccattgac tggtccgttc gagtttgagc cagacgtaat tcgatggtgg tggcgtgaga
     2341 tcaaaactca taagcattta ccttttaaac cgggcagcca ggcggcaata caccccggaa
     2401 gcatcacagg cctgtgcaaa cgtatggacg ccgacgctgt gccgaccaga ggcgagacta
     2461 ttgggaaaaa gactgcctcc tcagcatggg atcccgctac agttatgaga attttacgtg
     2521 accccagaat cgctggcttc gcagcggaag tgatttataa aaagaagcca gatggcacac
     2581 ccacaacaaa aattgagggg tacaggatcc agagagatcc tatcacttta aggccagttg
     2641 agttggattg tggaccaata attgaacccg ccgagtggta tgagcttcag gcctggttgg
     2701 atggaagagg tcgtggaaaa ggcctcagcc gtggacaggc catcctatct gctatggaca
     2761 aattgtattg cgaatgcggc gcggtgatga caagcaagcg aggcgaggaa tctatcaagg
     2821 acagctacag gtgtcgtaga cgtaaagttg ttgacccttc agcacctggc cagcatgagg
     2881 gtacgtgcaa tgtctccatg gcggcgttag acaagttcgt ggctgaacgt atatttaaca
     2941 aaattcgaca tgcagagggc gacgaggaga ctctagcgtt actatgggaa gccgcgcgtc
     3001 gatttgggaa gttaacggag gcgcctgaaa aaagcggaga gagagctaat ctcgtagccg
     3061 agcgtgctga tgcactgaat gcgctagagg agctgtatga ggaccgtgcc gccggggcat
     3121 acgatgggcc ggttggaagg aagcatttcc gtaaacagca agcggctctg acactgcgtc
     3181 agcaaggagc cgaagagcga ttagcggagc tcgaagccgc cgaggcgcct aaacttccac
     3241 tcgatcagtg gttccctgaa gacgcggacg ccgatccgac tgggcccaaa tcttggtggg
     3301 gtcgagcatc cgttgatgac aagcgtgtat ttgtgggtct atttgtcgac aagatcgtag
     3361 tgactaaatc aactacaggc cgtggccagg gcacgccaat cgagaaacgt gcgagcataa
     3421 catgggctaa accacctacg gacgacgatg aagacgatgc acaagatgga actgaggacg
     3481 ttgcagcata gggtaccgcg gccgcgaatt tccccgatcg ttcaaacatt tggcaataaa
     3541 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     3601 attacgttaa gcatgtaata tttaacatgt aatgcatgac gttatttatg agatgggttt
     3661 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatcgcgcg
     3721 caaacttgga taaattatcg cgcgcggtgt catctatgag gactagatcg caattcgccc
     3781 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3841 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3901 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3961 caaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4021 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4081 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4141 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4201 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4261 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4321 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4381 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     4441 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4501 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4561 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4621 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4681 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4741 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4801 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4861 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4921 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4981 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5041 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5101 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5161 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5221 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5281 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5341 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5401 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5461 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5521 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5581 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5641 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5701 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5761 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5821 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5881 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5941 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6001 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6061 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6121 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6181 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6241 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6301 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6361 cgc
//
