LOCUS       sequence-434013-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SWITCHER ready LoxP-EGFP reporter plasmid for cloning of a 
            customized CRISPR-inducible Cre-construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Schuster B, Dobiasovska I, Curcic J, Pajer P, Borna S, Bartunek P
  TITLE     SWITCHER, a CRISPR-inducible floxed wild-type Cre regulating CRISPR 
            activity.
  JOURNAL   Commun Biol. 2025 Jul 2;8(1):982. doi: 10.1038/s42003-025-08427-8.
  PUBMED    40596718
REFERENCE   2  (bases 1 to 6157)
  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: "2025-07-2"; volume: "8(1):982. doi"; pages: " 
            10.1038/s42003-025-08427-8"
COMMENT     Sequence Label: pSWITCHER-TF1-dsRED
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    11..80
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     enhancer        88..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        375..651
                     /label=chicken beta-actin promoter
     intron          652..879
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     primer_bind     892..911
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        892..910
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(955..988)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1022..2053
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPPEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     polyA_signal    2079..2213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2103..2122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2157..2176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(2338..2371)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2385..3062
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2565..2585)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2977..3000
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3233..3281
                     /note="synthetic polyadenylation signal"
     misc_feature    3295..3386
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3335..3354
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     repeat_region   3420..3560
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3420..3549
                     /label=AAV2 ITR
     rep_origin      3635..4090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3722..3741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3932..3953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4107..4126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4226..4248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4286..4304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4372..4476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4477..4545,4546..5337)
                     /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(4695..4714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5508..6096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5997..6016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtcacggt ctcgaagccg cggtgcgggt gccagggcgt gcccttgggc tccccgggcg
       61 cgtactccac ctcacccatc tggtacccgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
      181 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      241 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
      301 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      361 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      421 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      481 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
      541 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
      601 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      661 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      721 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      781 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
      841 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gtaatacgac
      901 tcactatagg gaagcttccg tatacaacct actacctcat aacatatgat atcgataact
      961 tcgtatagca tacattatac gaagttatcc gcgggcccgg gatccaccgg tcgccgccac
     1021 catgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
     1081 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
     1141 tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
     1201 ccggaaatgg tttcccccag aacctgaaga tgttcgcgat tatcttctat atcttcaggc
     1261 gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg
     1321 tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat
     1381 ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac
     1441 tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa
     1501 tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat
     1561 cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac
     1621 gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact
     1681 ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg
     1741 ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc
     1801 cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg
     1861 tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc
     1921 ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat
     1981 tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga
     2041 agatggcgat taagagctcg aattaattca tcgatgatga tccagacatg ataagataca
     2101 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2161 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2221 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaatgag
     2281 gtagtaggtt gtatacggag cttccagttt agccgtcgag ccagtttagc catcgacata
     2341 acttcgtata gcatacatta tacgaagtta tactagtgat atccatggcc tcctccgaga
     2401 acgtcatcac cgagttcatg cgcttcaagg tgcgcatgga gggcaccgtg aacggccacg
     2461 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggccacaac accgtgaagc
     2521 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttcc
     2581 agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac aagaagctgt
     2641 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggcga
     2701 ccgtgaccca ggactcctcc ctgcaggacg gctgcttcat ctacaaggtg aagttcatcg
     2761 gcgtgaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc tgggaggcct
     2821 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagacccac aaggccctga
     2881 agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg gccaagaagc
     2941 ccgtgcagct gcccggctac tactacgtgg acgccaagct ggacatcacc tcccacaacg
     3001 aggactacac catcgtggag cagtacgagc gcaccgaggg ccgccaccac ctgttcctgt
     3061 agcgggatat ccgggattcg tcagtagggt tgtaaaggtt tttcttttcc tgagaaaaca
     3121 accttttgtt ttctcaggtt ttgctttttg gcctttccct agctttaaaa aaaaaggacg
     3181 aaacaccacg cgtctggcct aactggccgg tacctggtat tggacaggcc gcaataaaat
     3241 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
     3301 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
     3361 aagtgcaggt gccagaacat ttctctggat cctgccatgt gtccactgag cgcggccgca
     3421 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3481 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3541 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3601 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     3661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     3721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3841 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3961 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4081 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4141 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4201 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4261 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4321 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4381 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4441 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4501 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4561 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4621 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4681 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4741 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4801 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4861 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4921 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4981 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5041 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5101 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5161 attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg
     5221 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5281 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5341 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5401 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5641 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5701 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5761 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5821 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5881 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5941 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6001 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6061 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6121 cggttcctgg ccttttgctg gccttttgct cacatgt
//
