LOCUS       sequence-171154-        8772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAc transposon with transformation marker for Tribolium 
            transgenesis; Drives EGFP expression in the eyes; contains Gal4_ 
            driven by Tribolium heatshock promoter; (Transformation plasmid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8772)
  TITLE     Bucher Lab plasmids
REFERENCE   2  (bases 1 to 8772)
  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-EGFP;Tc'hsp5'-Gal4Delta-3'UTR]
FEATURES             Location/Qualifiers
     source          1..8772
                     /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..3143)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2424..2426,2427..2429,2430..3140)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2469..2490)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2730..2749)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3077..3098
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3266..3387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3303..3322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3357..3376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4028..4372)
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             complement(4382..4822)
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     repeat_region   6336..6370
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6551..6567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6551..6567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6564..6586)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6575..6591
                     /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(6599..6605,6606..6623,6624..6629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6644..6665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6782..6799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6953..7541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7033..7052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7712..8503,8504..8572))
                     /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     8335..8354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8573..8677)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8745..8763
                     /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 accatggtga gcaagggcga ggagctgttc accggggtgg
     2461 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2521 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2581 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2641 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2701 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2761 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2821 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2881 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2941 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3001 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     3061 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3121 gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac
     3181 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3241 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3301 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3361 tttgtccaaa ctcatcaatg tatcttaaag cttatcgata cgcgtacggc gcgccaagct
     3421 taaggtgcac ggcccacgtg gccactagtg agcagccagt tgtttttttc tatattcctg
     3481 atgggcatac gaaggagagc ccgtttgcgc ttaatactgc caaggccacg actaatgcgg
     3541 taagtttttt attattttaa aatgaaacaa ctttttatct ttaaattaaa taatttatta
     3601 aaaattttta aataatctac aaaaatcagt ctttaaactt aatgcagtca ctatctactt
     3661 tattataact aaagtaactt tacgtactat tatttacaaa aataaactat cacaattaac
     3721 taagtaggaa cacaaaccga tcatctcgag ctctgtacat gtccgcggtc gacgtctctc
     3781 tagaagcttc tgaataagcc ctcgtaatat attttcatga agaatttagg tccaaaaaaa
     3841 agatgggcat taattctagt catttaaaaa attctataga tcagaggtta catggccaag
     3901 attgaaactt agaggagtat agttacataa aagaaggcaa aacgatgtat aaatgaaaga
     3961 aattgagatg gtgcacgatg cacagttgaa gtgaacttgc ggggtttttc agtatctacg
     4021 attcatttta ctcttttttt gggtttggtg gggtatcttc atcatcgaat agatagttat
     4081 atacatcatc cattgtagtg gtattaaaca tccctgtagt gattccaaac gcgttatacg
     4141 cagtttggtc cgtccaacca ggtgacagtg gttttgaatt attaccatca tcaattttac
     4201 tagccgtgat ttcattattc atgaagttat catgaacgtt agaggaggca attggttgtg
     4261 aaagcgcttg agaatttgtt tgagttgtta tgaggttcgg accgttgcta ctgttagtga
     4321 aagtgaagga caatgagcta tcagcaatat tcccactttg attaaaattg gcgaattccg
     4381 gcgatacagt caactgtctt tgacctttgt tactactctc ttccgatgat gatgtcgcac
     4441 ttattctatg ctgtctcaat gttagaggca tatcagtctc cactgaagcc aatctatctg
     4501 tgacggcatc tttattcaca ttatcttgta caaataatcc tgttaacaat gcttttatat
     4561 cctgtaaaga atccattttc aaaatcatgt caaggtcttc tcgaggaaaa atcagtagaa
     4621 atagctgttc cagtctttct agccttgatt ccacttctgt cagatgtgcc ctagtcagcg
     4681 gagacctttt ggttttggga gagtagcgac actcccagtt gttcttcaga cacttggcgc
     4741 acttcggttt ttctttggag cacttgagct ttttaagtcg gcaaatatcg catgcttgtt
     4801 cgatagaaga cagtagcttc atgggagacg tgtacctcta gatacacttt tcgatttact
     4861 ttgaattcac tagtaaataa ttcactcaac tttgttaaag tcgctttgaa aatttgcttt
     4921 gagtcgcttg cttagctttg ttgctttcgc ttcgcttgat tcaaattcac tgacaacgcg
     4981 ccgcgagacc gcgcttatat atgaaacggg gaattcgtct cgaacttcac cgaatgatct
     5041 cgaagtttcc attgatttgc gtttgaattt tctcgaattt ttatttttaa aacggccttt
     5101 tcggtattat tcatgctatt tccgtaaaga taaaggagaa tagtggaaac tggttgtgcc
     5161 ctaatttatt tcaacttcca aaataaaaac aatcattata tattaatgca tttaaaaaaa
     5221 aaacgttaca taacgaatga gttgttgact atttcatgtt tatttttaat ttggtaaagt
     5281 tcggcaatat tatgaatgag tagtgaattg ttttgatttc atatacatat gccaaagttg
     5341 gattgtcata gcaattttaa taaaacaatt ttcctatttt ttttcttttt aaacttatta
     5401 ataggtcttt aagtaaaatg ttttagatta atcctttttt ttacttttat cgtttataat
     5461 caattacttt tgcagaagat gcctaaataa tagtgattaa taataaataa agaacaattt
     5521 tttattgaag gttgggatcc atatataggg cccgggttat aattacctca ggtcgacgtc
     5581 ccatggccat tcgaattcgg ccggccgaat tcgaatggcc atgggacgtc gacctgaggt
     5641 aattataacc cgggccctat atatggatcc aattgcaatg atcatcatga cagatctgac
     5701 aatgttcagt gcagagactc ggctacgcct cgtggacttt gaagttgacc aacaatgttt
     5761 attcttacct ctaatagtcc tctgtggcaa ggtcaagatt ctgttagaag ccaatgaaga
     5821 acctggttgt tcaataacat tttgttcgtc taatatttca ctaccgcttg acgttggctg
     5881 cacttcatgt acctcatcta taaacgcttc ttctgtatcg ctctggacgt catcttcact
     5941 tacgtgatct gatatttcac tgtcagaatc ctcaccaaca agctcgtcat cgctttgcag
     6001 aagagcagag aggatatgct catcgtctaa agaactaccc attttattat atattagtca
     6061 cgatatctat aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat
     6121 aacaatataa ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc
     6181 attttgactc acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca
     6241 cgcctcacgg gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc
     6301 gctatttaga aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat
     6361 ctttctaggg ttaaaaaaga tttgcgcttt actcgaccta aactttaaac acgtcataga
     6421 atcttcgttt gacaaaaacc acattgtggc caagctgtgt gacgcgacgc gcgctaaaga
     6481 atggcaaacc aagtcgcgcg agcgtcgact ctagaggatc cccgggtacc gagctcgaat
     6541 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6601 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6661 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6721 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6781 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6841 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6901 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6961 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7021 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7081 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7141 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7201 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7261 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7321 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7381 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7441 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7501 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7561 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7621 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7681 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7741 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7801 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7861 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7921 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7981 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8041 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8101 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8161 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8221 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8281 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8341 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8401 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8461 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8521 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8581 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8641 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8701 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     8761 ggccctttcg tc
//
