LOCUS       sequence-170900-        9010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear reporter (histone-tagged EGFP) construct with the Tribolium 
            specific ribosomal protein subunit 3 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 9010)
  TITLE     Bucher Lab plasmids
REFERENCE   2  (bases 1 to 9010)
  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'rps3P-Tc'H2Av_EGFP]
FEATURES             Location/Qualifiers
     source          1..9010
                     /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(6330..6332,6333..6335,6336..7049)
                     /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(6330..6332,6333..6335,6336..7046)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6375..6396)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6636..6655)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6983..7004
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7078..7199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7089..7108)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7143..7162
                     /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 agatcttgtc aaaccacaaa cataaaaaat agatgacaga aattaacaaa attaccaaaa
     5281 aaactagaga atttatttat ttatttttgt ttttaattta tttttgccta tggtcttaca
     5341 ttgttttacg caacagttgc ataaattatt cggtccaaga tctttcccca agataatatt
     5401 ttacatttgg cgcaacttgt gtagaattac attgcgttta ttttctcaat tacggcgaaa
     5461 atatacgaca aaattgagat attttcaatt attcgtatgc aaaattgtct ggagaattga
     5521 ttggcgtcgt aaattcgtaa tagtttttca agttataaac cgaaatagat tttggctatt
     5581 tttgcatttg ttagcaataa taatattttt ggaaactacc agtagcagga caatgacctt
     5641 tttttaaacg atagatgatt aaaaagagtt ctgaacacat aaaagttcat aaaagttgtt
     5701 ggtttggttg gagctactca gataaaaaaa aaataatttt ttgaaggtaa aaaaactgaa
     5761 aaaagaacag tcttttcatt acgttattta cccgatatgc aagaaagttt tctctccaaa
     5821 aaattattca aataagccgc cttgtaagcg tcctctatca catcccttcc caattaggtc
     5881 gacttccgct cgccacacgt caaacctttt ccatttagaa cgtcaaaggc cggccctaaa
     5941 gaaaaatggc tggtggcaaa gcagggaaag attcgggcaa agctaaagcc aaggcggtgt
     6001 cgcgctcggc gcgtgccgga ctccagttcc ccgtggggag gatccaccgc cacttgaaga
     6061 atcgcactac gagccatgga cgcgtggggg ccaccgcggc tgtgtattcc gccgcgattt
     6121 tggagtattt gacagctgag gtcctggagt tggccgggaa cgccagtaag gacttgaagg
     6181 tgaagaggat cacccccagg cacttgcagc tggccatcag aggggatgag gagctggaca
     6241 gcctgatcaa ggccactatc gccggcggag gcgtcattcc acacatccac aagagtttga
     6301 tcgggaaaaa gggccactca cagcccgtca tggtgagcaa gggcgaggag ctgttcaccg
     6361 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     6421 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     6481 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     6541 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     6601 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     6661 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     6721 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     6781 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     6841 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     6901 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     6961 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     7021 ctctcggcat ggacgagctg tacaagtaag gcgcgccgta cgcgtatcga taagctttaa
     7081 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     7141 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     7201 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     7261 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctagag tcgcggccgc
     7321 cttaacgaaa taattaattg tttccattcg acacgttatt ggtgctattc gccgaccccc
     7381 aattgcacaa atgacttctc atagacgtag acaagggggg gatgtaggac cgtggtatca
     7441 aaaacgtcga taaattgaaa aggtccacga aaactttata ccgatcgctg aaaaacaaaa
     7501 tcaaacattg caaaaaccgt gttgaaatca atttcacctt aacgttgacc ttaaatactg
     7561 ataccccgaa gagccccctg ttcctaactg cgaagaacca atcattcttt gcaccatcaa
     7621 aacatgatta tctgaaatta taaaatttaa acaaaaacaa ttgacgtacg gatttactaa
     7681 caccgccatt tcgtaatcag cgagtctatg tccatcaaaa gtgttagaat ttggtggggc
     7741 tgactgaacc tgggttcgtc cctgtaaaag gtgatcatga tagccccttg cagggctttg
     7801 tgtgagaagc gcctctcgcc gcgcgacatt agagcctcat ggacctcaac tttaaaaatc
     7861 gactcgtaaa cctcgcgtct cttctccaaa tcgttcaatt tgtaacgctt taaagtctcc
     7921 gcctcctctt tctccgccag ttctttttgc tcggttaaaa gcttctcgac cgctttttgg
     7981 tacttccccc agaaattaaa cccctcgagc tccagccccg gcgttcgctc cagccaccgc
     8041 tggaccaaat cggtcaaaga cggctctttc tccgacttgg cgatttgctc gagcgctttt
     8101 tcgtcgtttc cgaacacttt cgagtaattc tggttgtatt tgacgcggtt ttcctggcgc
     8161 accccgagct tattttccag gagcctgaac tgcaggctct ggaacccgga agcgggccgc
     8221 agataacacc ggaagtccat gaaatcgagc ggtgtcatcg tttccagaat cattacttgg
     8281 tccaccaaaa cctacaattg cgacagtttt atctcgtgac cgcaatttgt tgcgaaattg
     8341 tgaaccttga ggatcaaaac aacacgattg aggcgtttga ggatttccaa agtttgcgac
     8401 tcttctaaaa cgtcgctgaa aatgttgcgg atcgagtcca gctcgtagat tatttgtttg
     8461 aaccataact catacgctgg aattacttgt tatagttaat tacggttaag tgggtttttt
     8521 accttgatgg gtcacaataa ataagtgctc gtcgtgcacg ggttggttat tttgctcgct
     8581 gagtagtctt tgggcttcta aaatcttgtc caacatcaag tattcaccgt aaagcatccc
     8641 acattcttcg ctcagttggt cgccttcttg ggcttcactt aatttacgat taattgtaaa
     8701 tggaataatt aaattaaatc atacttacga gggtctcagt gggcaactca ttacgaccgc
     8761 ggtaccgtcg actctagcgg taccccgatt gtttagcttg ttcagctgcg cttgtttatt
     8821 tgcttagctt tcgcttagcg acgtgttcac tttgcttgtt tgaattgaat tgtcgctccg
     8881 tagacgaagc gcctctattt atactccggc ggtcgagggt tcgaaatcga taagcttgga
     8941 tcctaattga attagctcta attgaattag tctctaattg aattagatcc ccgggcgagc
     9001 tcgaattaac
//
