LOCUS       sequence-171157-        7840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  UAS responder construct for misexpression in Tribolium; UAS drives 
            tGFP expression; (Transformation plasmid; Red eye marker).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7840)
  TITLE     Bucher Lab plasmids
REFERENCE   2  (bases 1 to 7840)
  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"
COMMENT     Sequence Label: pBac[3xP3-DsRed; UAS-Tc'hsp_p-tGFP-SV40]
FEATURES             Location/Qualifiers
     source          1..7840
                     /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"
     repeat_region   1099..1161
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      2418..2427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2424..2426,2427..2429,2430..3104)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(2607..2627)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2825..2844
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3019..3042
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3225..3346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3499..3593
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     regulatory      3782..3791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3788..4486
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     polyA_signal    4609..4730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5404..5438
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5619..5635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5619..5635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5632..5654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5643..5659
                     /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(5667..5673,5674..5691,5692..5697))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5712..5733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5850..5867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6021..6609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6101..6120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6780..7571,7572..7640))
                     /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     7403..7422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7641..7745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7813..7831
                     /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 ccagtgccaa gctttgttta aaaatataac
      421 aaaattgtga tcccacaaaa tgaagtgggg caaaatcaaa taattaatag tgtccgtaaa
      481 cttgttggtc ttcaactttt tgaggaacac gttggacggc aaatccgtga ctataacaca
      541 agttgattta ataattttag ccaacacgtc gggctgcgtg ttttttgccg acgcgtctgt
      601 gtacacgttg attaactggt cgattaaact gttgaaataa tttaattttt ggttcttctt
      661 taaatctgtg atgaaatttt ttaaaataac tttaaattct tcattggtaa aaaatgccac
      721 gttttgcaac ttgtgagggt ctaatatgag gtcaaactca gtaggagttt tatccaaaaa
      781 agaaaacatg attacgtctg tacacgaacg cgtattaacg cagagtgcaa agtataagag
      841 ggttaaaaaa tatattttac gcaccatata cgcatcgggt tgatatcgtt aatatggatc
      901 aatttgaaca gttgattaac gtgtctctgc tcaagtcttt gatcaaaacg caaatcgacg
      961 aaaatgtgtc ggacaatatc aagtcgatga gcgaaaaact aaaaaggcta gaatacgaca
     1021 atctcacaga cagcgttgag atatacggta ttcacgacag caggctgaat aataaaaaaa
     1081 ttagaaacta ttatttaacc ctagaaagat aatcatattg tgacgtacgt taaagataat
     1141 catgcgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta tttattaatt
     1201 tgaatagata ttaagtttta ttatatttac acttacatac taataataaa ttcaacaaac
     1261 aatttattta tgtttattta tttattaaaa aaaaacaaaa actcaaaatt tcttctataa
     1321 agtaacaaaa cttttaaaca ttctctcttt tacaaaaata aacttatttt gtactttaaa
     1381 aacagtcatg ttgtattata aaataagtaa ttagcttaac ttatacataa tagaaacaaa
     1441 ttatacttat tagtcagtca gaaacaactt tggcacatat caatattatg ctctcgacaa
     1501 ataacttttt tgcatttttt gcacgatgca tttgcctttc gccttatttt agaggggcag
     1561 taagtacagt aagtacgttt tttcattact ggctcttcag tactgtcatc tgatgtacca
     1621 ggcacttcat ttggcaaaat attagagata ttatcgcgca aatatctctt caaagtagga
     1681 gcttctaaac gcttacgcat aaacgatgac gtcaggctca tgtaaaggtt tctcataaat
     1741 tttttgcgac tttgaacctt ttctcccttg ctactgacat tatggctgta tataataaaa
     1801 gaatttatgc aggcaatgtt tatcattccg tacaataatg ccataggcca cctattcgtc
     1861 ttcctactgc aggtcatcac agaacacatt tggtctagcg tgtccactcc gcctttagtt
     1921 tgattataat acataaccat ttgcggttta ccggtacttt cgttgataga agcatcctca
     1981 tcacaagatg ataataagta taccatctta gctggcttcg gtttatatga gacgagagta
     2041 aggggtccgt caaaacaaaa catcgatgtt cccactggcc tggagcgact gtttttcagt
     2101 acttccggta tctcgcgttt gtttgatcgc acggttccca caatggttaa ttcgagctcg
     2161 cccggggatc taattcaatt agagactaat tcaattagag ctaattcaat taggatccaa
     2221 gcttatcgat ttcgaaccct cgaccgccgg agtataaata gaggcgcttc gtctacggag
     2281 cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta agcgaaagct aagcaaataa
     2341 acaagcgcag ctgaacaagc taaacaatcg gggtaccgct agagtcgacg gtaccgcggg
     2401 cccgggatcc accggtcgcc accatggtgc gctcctccaa gaacgtcatc aaggagttca
     2461 tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg
     2521 agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg
     2581 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
     2641 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
     2701 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     2761 ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg
     2821 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     2881 cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc
     2941 actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct
     3001 actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     3061 agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg cgactctaga
     3121 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3181 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3241 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3301 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaagc ttatcgatac
     3361 gcgtacggcg cgcctaggcc ggccgaattc gaatggccat gggacgtcga cctgaggtaa
     3421 ttataacccg ggccctatat atggatccaa ttgcaatgat catcatgaca gatctgcgcg
     3481 cgatcgatat ctgcaggtcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga
     3541 gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagac
     3601 tctagcgagc gccggagatc cgtttcatat ataagcgcgg tctcgcggcg cgttgtcagt
     3661 gaatttgaat caagcgaagc gaaagcaaca aagctaagca agcgactcaa agcaaatttt
     3721 caaagcgact ttaacaaagt tgagtgaatt atttactagt gaattcaaag tggtaccggt
     3781 cgccaccatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
     3841 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccgagca
     3901 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct
     3961 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
     4021 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
     4081 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
     4141 cggcgacttc aaggtgatgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
     4201 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgatctgga
     4261 tggcagcttc acccgcacct tcagcctgcg cgacggcggc tactacagct ccgtggtgga
     4321 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
     4381 gttcgccttc cgccgcgtgg aggaggatca cagcaacacc gagctgggca tcgtggagta
     4441 ccagcacgcc ttcaagaccc cggatgcaga tgccggtgaa gaataaagcg gccgcgactc
     4501 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4561 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4621 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4681 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta agcttggcgc
     4741 gcctaggccg gccgatctcg gatctgacaa tgttcagtgc agagactcgg ctacgcctcg
     4801 tggactttga agttgaccaa caatgtttat tcttacctct aatagtcctc tgtggcaagg
     4861 tcaagattct gttagaagcc aatgaagaac ctggttgttc aataacattt tgttcgtcta
     4921 atatttcact accgcttgac gttggctgca cttcatgtac ctcatctata aacgcttctt
     4981 ctgtatcgct ctggacgtca tcttcactta cgtgatctga tatttcactg tcagaatcct
     5041 caccaacaag ctcgtcatcg ctttgcagaa gagcagagag gatatgctca tcgtctaaag
     5101 aactacccat tttattatat attagtcacg atatctataa caagaaaata tatatataat
     5161 aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta
     5221 aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa
     5281 tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga
     5341 tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga
     5401 atgcatgcgt caattttacg cagactatct ttctagggtt aaaaaagatt tgcgctttac
     5461 tcgacctaaa ctttaaacac gtcatagaat cttcgtttga caaaaaccac attgtggcca
     5521 agctgtgtga cgcgacgcgc gctaaagaat ggcaaaccaa gtcgcgcgag cgtcgactct
     5581 agaggatccc cgggtaccga gctcgaattc gtaatcatgt catagctgtt tcctgtgtga
     5641 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5701 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5761 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5821 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5881 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6421 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6661 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6721 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6781 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6841 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6901 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6961 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7021 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7081 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7141 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7201 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7261 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7321 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7381 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7441 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7501 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7561 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7621 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7681 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7741 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7801 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
