LOCUS       sequence-268578-        8440 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 8440)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 8440)
  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..8440
                     /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
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWESSTERLYPRDGV
                     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   4997..5059
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5189..5206)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5189..5205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5198..5220)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5347..5802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5434..5453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5644..5665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5828..5932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5933..6001,6002..6793)
                     /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(6151..6170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6964..7552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7453..7472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7706..7723
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7840..7861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7876..7881,7882..7899,7900..7906)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7914..7930
                     /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     7919..7941
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7938..7954
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7938..7954
                     /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 gctgggagtc 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 aagccatacc aatggagctc acggggacag ccccccccca aagcccccag ggatgtaatt
     4621 acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg
     4681 ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc
     4741 acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg
     4801 gatacgggga aaaagaattc gataaaagtt ttgttacttt atagaagaaa ttttgagttt
     4861 ttgttttttt taataaataa ataaacataa ataaattgtt tgttgaattt attattagta
     4921 tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa acctcgatat
     4981 acagaccgat aaaacacatg cgtcaatttt acacatgatt atctttaacg tacgtcacaa
     5041 tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg agcatcttca
     5101 tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc tccacgggtt
     5161 caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     5221 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5281 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     5341 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5401 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5461 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5521 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5581 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5641 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5701 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5761 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     5821 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5941 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6001 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6061 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6121 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6241 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6481 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6541 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6781 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6841 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6901 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6961 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7021 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7081 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7141 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7201 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7261 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7321 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7381 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7441 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7501 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7561 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7621 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7681 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7741 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7801 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7861 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7921 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcccgccg
     7981 ggtaactcac ggggtatcca tgtccatttc tgcggcatcc agccaggata cccgtcctcg
     8041 ctgacgtaat atcccagcgc cgcaccgctg tcattaatct gcacaccggc acggcagttc
     8101 cggctgtcgc cggtattgtt cgggttgctg atgcgcttcg ggctgaccat ccggaactgt
     8161 gtccggaaaa gccgcgacga actggtatcc caggtggcct gaacgaacag ttcaccgtta
     8221 aaggcgtgca tggccacacc ttcccgaatc atcatggtaa acgtgcgttt tcgctcaacg
     8281 tcaatgcagc agcagtcatc ctcggcaaac tctttccatg ccgcttcaac ctcgcgggaa
     8341 aaggcacggg cttcttcctc cccgatgccc agatagcgcc agcttgggcg atgactgagc
     8401 cggaaaaaag acccgacgat atgatcctga tgcagctaga
//
