LOCUS       sequence-256382-        8439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for constitutive expression of tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8439)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 8439)
  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: BII-ChPtW-tdT
FEATURES             Location/Qualifiers
     source          1..8439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(308..324)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1875..1928
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1968..3398
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    3416..4004
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3469..3489)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3887..3898)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4084..4218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4108..4127)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4162..4181
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4996..5058
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5188..5205)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5188..5204)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5197..5219)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5346..5801
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5433..5452)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5643..5664
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5827..5931
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5932..6000,6001..6792)
                     /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(6150..6169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6963..7551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7452..7471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7705..7722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7839..7860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7875..7880,7881..7898,7899..7905)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7913..7929
                     /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     7918..7940
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7937..7953
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7937..7953
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat
      361 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
      421 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
      481 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
      541 ggatacgggg aaaaggcctc caaggcctgc aggagtcaat gggaaaaacc cattggagcc
      601 aagtacactg actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag
      661 ggggtgagtc aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc
      721 caatgggttt tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt
      781 actgacatgt atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt
      841 caataggggt gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga
      901 ctttccattg ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc
      961 ccattattgg cacatacata aggtcaatag gggtgactag tggagaagag catgcttgag
     1021 ggctgagtgc ccctcagtgg gcagagagca catggcccac agtccctgag aagttggggg
     1081 gaggggtggg caattgaact ggtgcctaga gaaggtgggg cttgggtaaa ctgggaaagt
     1141 gatgtggtgt actggctcca cctttttccc cagggtgggg gagaaccata tataagtgca
     1201 gtagtctctg tgaacattcg gtttagtgaa cctgcaggtc ggccgccacg accggtgcca
     1261 ccatgactga atacaaacca actgttcgcc tggcaactcg tgatgatgtt ccacgtgcag
     1321 ttcgcaccct ggctgctgca tttgctgact accctgcaac ccgtcacact gtggacccag
     1381 accgccacat tgaacgtgtg actgaactgc aggagctgtt cctgacccgt gtgggcctgg
     1441 acattggcaa agtgtgggtg gcagatgatg gtgctgctgt ggcagtgtgg accacccctg
     1501 aatctgttga agctggtgca gtgtttgctg agattggccc acgcatggca gaactgtctg
     1561 gcagccgcct ggcagcacaa cagcagatgg aaggtctgct ggcaccacac cgcccaaaag
     1621 aacctgcttg gttcctggca actgtgggtg tgagccctga ccaccagggt aagggcctgg
     1681 gctctgcagt ggtgctgcct ggtgtggaag cagctgaacg tgcaggtgtg cctgctttcc
     1741 tggagacctc agctccacgc aacctgcctt tctatgaacg cctgggcttc actgtgactg
     1801 ctgatgtgga agtgccagaa ggcccacgca cttggtgcat gactcgcaaa ccaggtgcta
     1861 gccttggctc cggcgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     1921 ccggcccact cggctacccc tacgacgtgc ccgactacgc cagcggcatg gtgagcaagg
     1981 gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     2041 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2101 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     2161 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     2221 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     2281 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     2341 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2401 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     2461 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     2521 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     2581 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     2641 tcctggggca tggcaccggc agcaccggca gcggcagctc cggcaccgcc tcctccgagg
     2701 acaacaacat ggccgtcatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca
     2761 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2821 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
     2881 ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac atccccgatt
     2941 acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     3001 gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg atctacaagg
     3061 tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg
     3121 gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc
     3181 accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag accatctaca
     3241 tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag ctggacatca
     3301 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc
     3361 acctgttcct gtacggcatg gacgagctgt acaagtaaga gttcgcgtct ggaacaatca
     3421 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3481 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3541 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3601 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3661 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3721 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3781 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3841 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3901 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3961 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgg
     4021 ctaagcactt cgtggccgtc gatcgtttaa agggaggtag tgagtcgaga tatcgatact
     4081 agtgatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4141 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4201 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4261 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     4321 gatcctctag agggacagcc cccccccaaa gcccccaggg atgtaattac gtccctcccc
     4381 cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct ccccccgcat
     4441 ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac gggatcgctt
     4501 tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga tacggggaaa
     4561 agccatacca atggagctca cggggacagc ccccccccaa agcccccagg gatgtaatta
     4621 cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc ggtccggcgc
     4681 tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca
     4741 cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac acctgggggg
     4801 atacggggaa aaagaattcg ataaaagttt tgttacttta tagaagaaat tttgagtttt
     4861 tgtttttttt aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat
     4921 gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata
     4981 cagaccgata aaacacatgc gtcaatttta cacatgatta tctttaacgt acgtcacaat
     5041 atgattatct ttctagggtt aatctagctg cgtgttctgc agcgtgtcga gcatcttcat
     5101 ctgctccatc acgctgtaaa acacatttgc accgcgagtc tgcccgtcct ccacgggttc
     5161 aaaaacgtga atgaacgagg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5221 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5281 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5341 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5401 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5461 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5521 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5581 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5641 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5701 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5761 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     5821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5941 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6001 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6061 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6121 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6181 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6241 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6301 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6361 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6421 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6481 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6541 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6601 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6661 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6721 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6781 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6841 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6901 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6961 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7021 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7081 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     7141 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7201 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7261 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7321 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7381 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7441 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7501 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7561 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7621 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7681 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7741 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7801 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7861 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7921 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcccgccgg
     7981 gtaactcacg gggtatccat gtccatttct gcggcatcca gccaggatac ccgtcctcgc
     8041 tgacgtaata tcccagcgcc gcaccgctgt cattaatctg cacaccggca cggcagttcc
     8101 ggctgtcgcc ggtattgttc gggttgctga tgcgcttcgg gctgaccatc cggaactgtg
     8161 tccggaaaag ccgcgacgaa ctggtatccc aggtggcctg aacgaacagt tcaccgttaa
     8221 aggcgtgcat ggccacacct tcccgaatca tcatggtaaa cgtgcgtttt cgctcaacgt
     8281 caatgcagca gcagtcatcc tcggcaaact ctttccatgc cgcttcaacc tcgcgggaaa
     8341 aggcacgggc ttcttcctcc ccgatgccca gatagcgcca gcttgggcga tgactgagcc
     8401 ggaaaaaaga cccgacgata tgatcctgat gcagctaga
//
