LOCUS       sequence-170877-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAc transposon with transformation marker for Tribolium 
            transgenesis; drives ECFP (blue fluorescent) expression in the eyes 
            (much stronger than usual 3XP3); (Transformation plasmid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  TITLE     Bucher Lab plasmids
REFERENCE   2  (bases 1 to 6463)
  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[6xP3-Tc-hsp-ECFP-SV40]
FEATURES             Location/Qualifiers
     source          1..6463
                     /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      2409..2418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2415..2417,2418..2420,2421..3134)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2460..2481)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2721..2740)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3068..3089
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3257..3378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3294..3313)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3348..3367
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4027..4061
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4242..4258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4242..4258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4255..4277)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4266..4282
                     /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(4290..4296,4297..4314,4315..4320))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4335..4356
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4473..4490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4644..5232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4724..4743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5403..6194,6195..6263))
                     /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     6026..6045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6264..6368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6436..6454
                     /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 caatggtcga cgatctaatt
     2161 caattagaga ctaattcaat tagagctaat tcaattaaga tctaattcaa ttagagacta
     2221 attcaattag agctaattca attaatccgt ttcatatata agcgcggtct cgcggcgcgt
     2281 tgtcagtgaa tttgaatcaa gcgaagcgaa agcaacaaag ctaagcaagc gactcaaagc
     2341 aaattttcaa agcgacttta acaaagttga gtgaattatt tactagtgaa ttcaaagtgg
     2401 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2461 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2521 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2581 tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc
     2641 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2701 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2761 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2821 acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg
     2881 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca
     2941 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3001 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3061 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3121 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     3181 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3241 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3301 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3361 actcatcaat gtatcttaaa gctgatctga caatgttcag tgcagagact cggctacgcc
     3421 tcgtggactt tgaagttgac caacaatgtt tattcttacc tctaatagtc ctctgtggca
     3481 aggtcaagat tctgttagaa gccaatgaag aacctggttg ttcaataaca ttttgttcgt
     3541 ctaatatttc actaccgctt gacgttggct gcacttcatg tacctcatct ataaacgctt
     3601 cttctgtatc gctctggacg tcatcttcac ttacgtgatc tgatatttca ctgtcagaat
     3661 cctcaccaac aagctcgtca tcgctttgca gaagagcaga gaggatatgc tcatcgtcta
     3721 aagaactacc cattttatta tatattagtc acgatatcta taacaagaaa atatatatat
     3781 aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc
     3841 ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca
     3901 aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg
     3961 agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca
     4021 agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaaaaaag atttgcgctt
     4081 tactcgacct aaactttaaa cacgtcatag aatcttcgtt tgacaaaaac cacattgtgg
     4141 ccaagctgtg tgacgcgacg cgcgctaaag aatggcaaac caagtcgcgc gagcgtcgac
     4201 tctagaggat ccccgggtac cgagctcgaa ttcgtaatca tgtcatagct gtttcctgtg
     4261 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4321 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4381 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4441 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4501 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4561 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4621 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4681 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4741 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4801 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4861 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4921 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4981 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5041 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5101 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5161 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5221 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5281 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5341 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5401 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5461 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5521 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5581 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5641 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5701 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5761 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5821 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5881 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5941 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6001 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6061 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6121 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6181 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6241 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6301 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6361 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6421 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
