LOCUS       sequence-429021-        7190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tol2 based UAS modified with a IL2RGTC binding site upstream of it, 
            driving GPHN-FingR fused to mCherry, IL2RGTC zinc finger, and 
            KRAB(A) repressor domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7190)
  TITLE     Bonkowsky lab FingR plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7190)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTol2-ziUAS:GPHN.FingR-mCherry-ZFI (IL2RGTC)-K...
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     383..399
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    419..439
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     441..460
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        441..459
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(512..616)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1228..1244)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1281..1299)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1299..1323
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1597..2304
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1742..1760)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2010..2029
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2204..2227
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2745..2870
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     intron          2913..3045
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3028..3047
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(3107..3127)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(3186..3210)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3212..3346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3409..3429)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3437..3453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3469..3603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(4042..4229,4230..4241)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4275..4291)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4328..4346)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4367..4383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4380..4402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4391..4407
                     /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(4415..4421,4422..4439,4440..4445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4460..4481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4598..4615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4769..5357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4849..4868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5528..6319,6320..6388))
                     /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     6151..6170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6389..6493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6656..6677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(7026..7037,7038..7190,1..10)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 aaagcaagaa agaaaactag agattcttgt ttaagctttt aatctcaaaa aacattaaat
       61 gaaatgcata caaggtttta tcctgcttta gaactgtttg tatttaatta tcaaactata
      121 agacagacaa tctaatgcca gtacacgcta ctcaaagttg taaaacctca gatttaactt
      181 cagtagaagc tgattctcaa aattgttagt gtcaagccta gctcttttgg ggctgaaaag
      241 caatcctgca gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag gtgtattagt
      301 cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac
      361 acaggccaga tgggccctcg agcaggaaac agctatgacc atgattacgc caagctatca
      421 actttgtata gaaaagttgg taatacgact cactataggg cgaattgggt accaagggaa
      481 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      541 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      601 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg
      661 actagtcttc cacagagtac tagttattaa tagtaatcaa ttacggggtc attagttcat
      721 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      781 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      841 gggactttcc attgacgtca atgggtggac tatttacggt aagcttaggc ctccaaggcg
      901 gagtactgtc ctccgggctg gcggagtact gtcctccggc aaggtcggag tactgtcctc
      961 cgacactaga ggtcggagta ctgtcctccg acgcaaggcg gagtactgtc ctccgggctg
     1021 cggagtactg tcctccggca aggtcggagt actgtcctcc gacactagag gtcggagtac
     1081 tgtcctccga cgcaaggcgg agtactgtcc tccgggctgg cggagtactg tcctccggca
     1141 agggtcgact ctagagggta tataatggat cccatcgcgt ctcagcctca ctttgagctc
     1201 ctccacacga attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg
     1261 tggagctcca gcttttgttc cctttagtga gggttaattc aagtttgtac aaaaaagcag
     1321 gctaccatgc tcgaagtcaa ggaagcatca ccaaccagca tccagatcag ctggggcaag
     1381 tacaaggtca tggttcgcta ctaccgcatc acctacggtg aaactggtgg caatagccct
     1441 gtccaggaat tcaccgtgcc tggcagcaag tccactgcta ccatcagcag cctgaaacct
     1501 ggtgtcgact ataccatcac ggtgtacgcc gtcacgatcg accactggaa ctaccaggac
     1561 ccgatcccga tctccatcaa ctaccgcacc ggatccatgg tgagcaaggg cgaggaggac
     1621 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     1681 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     1741 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtcccct
     1801 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     1861 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     1921 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     1981 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     2041 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     2101 aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac ctacaaggcc
     2161 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagctgga catcacctcc
     2221 cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg ccactccacc
     2281 ggcggcatgg acgagctgta caagtggcca gatactgcga cctccctagc aaactggggc
     2341 acagatctag atttcagtgc cagatttgca tgcgcaactt tagccgcaaa agcaccctga
     2401 ccgatcatat tcgcacccat accggcgaaa aaccgtttgc gtgcgatatt tgcggccgca
     2461 aatttgcggc gcgcagcacc cgcaccaccc ataccaaaat tcataccggc agccagaaac
     2521 cgtttcagtg ccgcatttgc atgcgcaact ttagccgcag cgatagcctg agcaaacata
     2581 ttcgcaccca taccggcgaa aaaccgtttg cgtgcgatat ttgcggccgc aaatttgcgc
     2641 agcgcagcaa cctgaaagtg cataccaaaa ttcatctgcg cggcagccag ctgatcgatg
     2701 gtgtcgacgc tcctgaacaa cgtgaaggtg cttctcaagt ttctgtttct gttacttttg
     2761 aagatgttgc tgttcttttt actcgtgatg aatggaaaaa acttgatctt tctcaacgtt
     2821 ctctttatcg tgaagttatg cttgaaaatt attctaatct tgcttctatg gcttagcgta
     2881 caagtaaagc ggccccggac tcgaggccgc aggtaagtat caaggttaca agacaggttt
     2941 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     3001 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc gacaatcaac
     3061 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3121 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3181 tcattaccca gctttcttgt acaaagtggg ggatccagac atgataagat acattgatga
     3241 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3301 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3361 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcca actttattat
     3421 acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga tccagacatg
     3481 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3541 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3601 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3661 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagatcag
     3721 atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg actgtgctgc
     3781 tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg actgttataa
     3841 tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga catgacgtca
     3901 cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa cgattttcat
     3961 aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat tggtttggta
     4021 atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt gactgtaaat
     4081 aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa
     4141 aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata atacttaagt
     4201 acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac cccctacctt
     4261 cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg agctccagct
     4321 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     4381 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4441 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4501 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5401 acgaaaactc acgttaaggc attttggtca tgagattatc aaaaaggatc ttcacctaga
     5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6481 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     6541 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6601 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6661 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6721 ctatcagggc gatggcccac gctggctaag aactcatcag cctccccggt ccatctaccc
     6781 acgtaccaat gcaccaattg gccacaatga cggctactac atggtgccat tccttcctct
     6841 ttataggaat ggagactacc tcctgtccaa caatgctctt ggatacgagt acgcctacct
     6901 gttggaccca ggtcattgca caacaccaga aatgccctct gatctgcaaa agacgtgaat
     6961 atctgttcag acacccatat ccactctgtt ccacacaggt cagaggtttg tccaggagtt
     7021 cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac aagtacttag
     7081 ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt gagtaaaagt
     7141 aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta
//
