LOCUS       sequence-171155-        9257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear reporter (histone-tagged EGFP) construct with Tribolium 
            specific polyubiquitin promoter (driving ubiquitous expression); 
            (Transformation plasmid; Black eye marker).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9257)
  TITLE     Bucher Lab plasmids
REFERENCE   2  (bases 1 to 9257)
  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-gTc'v;Tc'puP-Tc'H2Av_EGFP]
FEATURES             Location/Qualifiers
     source          1..9257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   985..1047
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1751..1768)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1751..1767)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1760..1782)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1976..1995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2095..2117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2155..2173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2241..2345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2346..2414,2415..3206)
                     /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(2564..2583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3377..3965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3866..3885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4119..4136
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4253..4274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4289..4294,4295..4312,4313..4319)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4327..4343
                     /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     4332..4354
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4351..4367
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4351..4367
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4548..4582
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             join(6577..6579,6580..6582,6583..7296)
                     /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(6577..6579,6580..6582,6583..7293)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6622..6643)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6883..6902)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7230..7251
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7325..7446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7336..7355)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7390..7409
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cattgtggga accgtgcgat caaacaaacg cgagataccg gaagtactga aaaacagtcg
       61 ctccaggcca gtgggaacat cgatgttttg ttttgacgga ccccttactc tcgtctcata
      121 taaaccgaag ccagctaaga tggtatactt attatcatct tgtgatgagg atgcttctat
      181 caacgaaagt accggtaaac cgcaaatggt tatgtattat aatcaaacta aaggcggagt
      241 ggacacgcta gaccaaatgt gttctgtgat gacctgcagt aggaagacga ataggtggcc
      301 tatggcatta ttgtacggaa tgataaacat tgcctgcata aattctttta ttatatacag
      361 ccataatgtc agtagcaagg gagaaaaggt tcaaagtcgc aaaaaattta tgagaaacct
      421 ttacatgagc ctgacgtcat cgtttatgcg taagcgttta gaagctccta ctttgaagag
      481 atatttgcgc gataatatct ctaatatttt gccaaatgaa gtgcctggta catcagatga
      541 cagtactgaa gagccagtaa tgaaaaaacg tacttactgt acttactgcc cctctaaaat
      601 aaggcgaaag gcaaatgcat cgtgcaaaaa atgcaaaaaa gttatttgtc gagagcataa
      661 tattgatatg tgccaaagtt gtttctgact gactaataag tataatttgt ttctattatg
      721 tataagttaa gctaattact tattttataa tacaacatga ctgtttttaa agtacaaaat
      781 aagtttattt ttgtaaaaga gagaatgttt aaaagttttg ttactttata gaagaaattt
      841 tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg ttgaatttat
      901 tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta ataaataaac
      961 ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat ctttaacgta
     1021 cgtcacaata tgattatctt tctagggtta aataatagtt tctaattttt ttattattca
     1081 gcctgctgtc gtgaataccg tatatctcaa cgctgtctgt gagattgtcg tattctagcc
     1141 tttttagttt ttcgctcatc gacttgatat tgtccgacac attttcgtcg atttgcgttt
     1201 tgatcaaaga cttgagcaga gacacgttaa tcaactgttc aaattgatcc atattaacga
     1261 tatcaacccg atgcgtatat ggtgcgtaaa atatattttt taaccctctt atactttgca
     1321 ctctgcgtta atacgcgttc gtgtacagac gtaatcatgt tttctttttt ggataaaact
     1381 cctactgagt ttgacctcat attagaccct cacaagttgc aaaacgtggc attttttacc
     1441 aatgaagaat ttaaagttat tttaaaaaat ttcatcacag atttaaagaa gaaccaaaaa
     1501 ttaaattatt tcaacagttt aatcgaccag ttaatcaacg tgtacacaga cgcgtcggca
     1561 aaaaacacgc agcccgacgt gttggctaaa attattaaat caacttgtgt tatagtcacg
     1621 gatttgccgt ccaacgtgtt cctcaaaaag ttgaagacca acaagtttac ggacactatt
     1681 aattatttga ttttgcccca cttcattttg tgggatcaca attttgttat atttttaaac
     1741 aaagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1801 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1861 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     1921 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     1981 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2041 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2101 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     2161 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2221 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2281 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2341 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2401 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2461 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2521 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2581 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2641 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2701 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2761 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2821 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2881 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2941 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3001 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3061 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3121 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3181 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3241 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3301 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3361 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3961 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4021 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4081 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4141 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4201 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     4261 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     4321 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt
     4381 cgagctcggt acccggggat cctctagagt cgacgctcgc gcgacttggt ttgccattct
     4441 ttagcgcgcg tcgcgtcaca cagcttggcc acaatgtggt ttttgtcaaa cgaagattct
     4501 atgacgtgtt taaagtttag gtcgagtaaa gcgcaaatct tttttaaccc tagaaagata
     4561 gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg
     4621 aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc
     4681 ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac
     4741 gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt
     4801 tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata gatatcgtga
     4861 ctaatatata ataaaatggg tagttcttta gacgatgagc atatcctctc tgctcttctg
     4921 caaagcgatg acgagcttgt tggtgaggat tctgacagtg aaatatcaga tcacgtaagt
     4981 gaagatgacg tccagagcga tacagaagaa gcgtttatag atgaggtaca tgaagtgcag
     5041 ccaacgtcaa gcggtagtga aatattagac gaacaaaatg ttattgaaca accaggttct
     5101 tcattggctt ctaacagaat cttgaccttg ccacagagga ctattagagg taagaataaa
     5161 cattgttggt caacttcaaa gtccacgagg cgtagccgag tctctgcact gaacattgtc
     5221 agatcttgta cttttctttg tcccaaatga cccttttaca taatttatta ttgtcgtccg
     5281 tatttacatg ataaacaaga ctctagaaca acaacgataa ggataattct ttgtaaacaa
     5341 attaattcgt ttcaaaattg aacagacgaa tttggcattc catgcaggaa ttaatgggtg
     5401 gtttatttga aacgtttgaa aagcataaat tgggaaataa attaaaattt gaaaaataaa
     5461 aaagggcgta attccaaaac aaagaaaaca gctgatcgat aaagctgacg acgtagataa
     5521 aaatcagctg acatgtcacc aacaactgct ggccaaagtc cctaaaatct tgttcaatca
     5581 aaagcaattt tcttaatacc tgaaacattg tttagaccat gattaaccga attggcatta
     5641 aactaacagc gactaacgtt tgtgcataaa tttttgatta aagaggacgt tttttggata
     5701 attactcgaa aattctagaa agtgtacttt gggttcgaga ttaaaaagga gtgggggaag
     5761 ccgtccgtat aaaagcggtt ccacagctgc attcgccata ttgtctgagt aaaagtggtg
     5821 aaatacaaaa gcgatttaat taaaattgtg aattttccta gccgtgaagg tatgtatttt
     5881 aatttcatca tgaattattc gattgcaaga agcgcagaaa gtcgcaattt tgtgaaattt
     5941 ttcgcatgac gcaataacat ttgtttgaaa aaacatcgcc ctagtgacgt gtttttttgt
     6001 ttaaatttta acaaaattaa ttttgtaaat ttgtcgatct gttcaaaacc aaaccacata
     6061 gtgagtaatg tgttttttcg gtttattcag tgcgtaattc tttgttcaac gaaatccaat
     6121 atggcggcct ggaaatttgc tttgtcacaa aagtaatttt ttgtgtcgtt gcagggccgg
     6181 ccctaaagaa aaatggctgg tggcaaagca gggaaagatt cgggcaaagc taaagccaag
     6241 gcggtgtcgc gctcggcgcg tgccggactc cagttccccg tggggaggat ccaccgccac
     6301 ttgaagaatc gcactacgag ccatggacgc gtgggggcca ccgcggctgt gtattccgcc
     6361 gcgattttgg agtatttgac agctgaggtc ctggagttgg ccgggaacgc cagtaaggac
     6421 ttgaaggtga agaggatcac ccccaggcac ttgcagctgg ccatcagagg ggatgaggag
     6481 ctggacagcc tgatcaaggc cactatcgcc ggcggaggcg tcattccaca catccacaag
     6541 agtttgatcg ggaaaaaggg ccactcacag cccgtcatgg tgagcaaggg cgaggagctg
     6601 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     6661 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     6721 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     6781 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     6841 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     6901 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     6961 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     7021 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     7081 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     7141 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     7201 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     7261 gggatcactc tcggcatgga cgagctgtac aagtaaggcg cgccgtacgc gtatcgataa
     7321 gctttaagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     7381 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     7441 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     7501 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tctagagtcg
     7561 cggccgcctt aacgaaataa ttaattgttt ccattcgaca cgttattggt gctattcgcc
     7621 gacccccaat tgcacaaatg acttctcata gacgtagaca agggggggat gtaggaccgt
     7681 ggtatcaaaa acgtcgataa attgaaaagg tccacgaaaa ctttataccg atcgctgaaa
     7741 aacaaaatca aacattgcaa aaaccgtgtt gaaatcaatt tcaccttaac gttgacctta
     7801 aatactgata ccccgaagag ccccctgttc ctaactgcga agaaccaatc attctttgca
     7861 ccatcaaaac atgattatct gaaattataa aatttaaaca aaaacaattg acgtacggat
     7921 ttactaacac cgccatttcg taatcagcga gtctatgtcc atcaaaagtg ttagaatttg
     7981 gtggggctga ctgaacctgg gttcgtccct gtaaaaggtg atcatgatag ccccttgcag
     8041 ggctttgtgt gagaagcgcc tctcgccgcg cgacattaga gcctcatgga cctcaacttt
     8101 aaaaatcgac tcgtaaacct cgcgtctctt ctccaaatcg ttcaatttgt aacgctttaa
     8161 agtctccgcc tcctctttct ccgccagttc tttttgctcg gttaaaagct tctcgaccgc
     8221 tttttggtac ttcccccaga aattaaaccc ctcgagctcc agccccggcg ttcgctccag
     8281 ccaccgctgg accaaatcgg tcaaagacgg ctctttctcc gacttggcga tttgctcgag
     8341 cgctttttcg tcgtttccga acactttcga gtaattctgg ttgtatttga cgcggttttc
     8401 ctggcgcacc ccgagcttat tttccaggag cctgaactgc aggctctgga acccggaagc
     8461 gggccgcaga taacaccgga agtccatgaa atcgagcggt gtcatcgttt ccagaatcat
     8521 tacttggtcc accaaaacct acaattgcga cagttttatc tcgtgaccgc aatttgttgc
     8581 gaaattgtga accttgagga tcaaaacaac acgattgagg cgtttgagga tttccaaagt
     8641 ttgcgactct tctaaaacgt cgctgaaaat gttgcggatc gagtccagct cgtagattat
     8701 ttgtttgaac cataactcat acgctggaat tacttgttat agttaattac ggttaagtgg
     8761 gttttttacc ttgatgggtc acaataaata agtgctcgtc gtgcacgggt tggttatttt
     8821 gctcgctgag tagtctttgg gcttctaaaa tcttgtccaa catcaagtat tcaccgtaaa
     8881 gcatcccaca ttcttcgctc agttggtcgc cttcttgggc ttcacttaat ttacgattaa
     8941 ttgtaaatgg aataattaaa ttaaatcata cttacgaggg tctcagtggg caactcatta
     9001 cgaccgcggt accgtcgact ctagcggtac cccgattgtt tagcttgttc agctgcgctt
     9061 gtttatttgc ttagctttcg cttagcgacg tgttcacttt gcttgtttga attgaattgt
     9121 cgctccgtag acgaagcgcc tctatttata ctccggcggt cgagggttcg aaatcgataa
     9181 gcttggatcc taattgaatt agctctaatt gaattagtct ctaattgaat tagatccccg
     9241 ggcgagctcg aattaac
//
