LOCUS       sequence-274028-        7808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG tdTomato piggyBac transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7808)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7808)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: PB CAG-tdTomato
FEATURES             Location/Qualifiers
     source          1..7808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        189..568
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        570..847
                     /label=chicken beta-actin promoter
     intron          848..1864
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1788..1810
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1872..1891
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1927..1936
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1933..3363
                     /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    3403..3991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3456..3476)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3874..3885)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4030..4047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4036..4260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4614..4648
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4735..4754)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4736..4754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4761..4778)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4761..4777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4770..4792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4919..5374)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5056..5077)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5268..5287
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5401..5505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5506..5574,5575..6366)
                     /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(5724..5743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6537..7125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7026..7045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7279..7296
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7413..7434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7449..7454,7455..7472,7473..7479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7487..7503
                     /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     7492..7514
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7511..7527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7511..7527
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7546..7566
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7548..7566
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   7630..7692
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ttatttatta aaaaaaaaca aaaactcaaa atttcttcta taaagtaaca aaacttttaa
       61 acattctctc ttttacaaaa ataaacttat tttgtacttt aaaaacagtc atgttgtatt
      121 ataaaataag taattagctt aacctataca taatagaaac aaattatact tattaaccgg
      181 tcatggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      241 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      301 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      361 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      421 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      481 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      541 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      601 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      661 tgcagcgatg ggggcggggg gggggggggg gcgcgcgccr gggggggggg gggggggggg
      721 gggggggggs ggggsgrggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
      781 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      841 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      901 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      961 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
     1021 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
     1081 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
     1141 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
     1201 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
     1261 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
     1321 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
     1381 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
     1441 cggggcgggg ccgcctcggg ccggggrggg ctcgggggag gggcgcggcg gcccccggag
     1501 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     1561 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
     1621 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     1681 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1741 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1801 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1861 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgct
     1921 agcgccgcca ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
     1981 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     2041 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     2101 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     2161 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     2221 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     2281 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     2341 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     2401 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     2461 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     2521 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     2581 cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc
     2641 agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc
     2701 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     2761 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     2821 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     2881 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     2941 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     3001 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     3061 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     3121 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     3181 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     3241 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     3301 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     3361 taataattaa ttaatgatca tacagtgctc tgagtacgct ataatcaacc tctggattac
     3421 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3481 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3541 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3601 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3661 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3721 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3781 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     3841 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3901 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3961 agtcggatct ccctttgggc cgcctccccg caggatccac tagagcggcc gcctagagct
     4021 cgctatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4081 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4141 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4201 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4261 cttctgaggc ggaaagaacc agctggggct cgactagagc ttgcggaacc cttaagcttt
     4321 tttattatat attagtcacg atatctataa caagaaaata tatatataat aagttatcac
     4381 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
     4441 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
     4501 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
     4561 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
     4621 caattttacg cagactatct ttctagggtt aaaaaagatt tgcgctttac tcgacctaaa
     4681 ctttaaacac gtcatagaat cttcgtttga caaaaaccac attgtgttgg gtcgccctat
     4741 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4801 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4861 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcaa
     4921 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4981 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5041 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5101 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5161 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5221 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5281 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5341 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     5401 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5461 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5521 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5581 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5641 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5701 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5761 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5821 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5881 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5941 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6001 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6061 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6661 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7141 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7201 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7261 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7321 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     7381 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7441 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7501 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     7561 taaagggaac ctcccccggt attcacgaca gcaggctgaa taataaaaaa attagaaact
     7621 attatttaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa
     7681 aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat
     7741 attaagtttt attatattta cacttacata ctaataataa attcaacaaa caatttattt
     7801 atgtttat
//
