LOCUS       sequence-316225-        7991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PB-CA tdTomato piggyBac transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7991)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7991)
  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            "Unpublished"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-CA tdTomato
FEATURES             Location/Qualifiers
     source          1..7991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1739..1748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1745..3175
                     /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    3215..3803
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3268..3288)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3686..3697)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3842..3859)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3848..4072
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4497..4531
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(4680..4698)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4719..4735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4719..4735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4732..4754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4743..4759
                     /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(4767..4773,4774..4791,4792..4797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4812..4833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4950..4967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5121..5709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5201..5220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5880..6671,6672..6740))
                     /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     6503..6522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6741..6845)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6871..7326)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7008..7029)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7220..7239
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7453..7475
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7467..7484
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7468..7484
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7491..7510
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7491..7509
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   7576..7638
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc cggggggggs gggggggggs ggggggsggg
      541 gsggggsgrg gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggrg ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg ctagcgccgc
     1741 caccatggtg agcaagggcg aggaggtcat caaagagttc atgcgcttca aggtgcgcat
     1801 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1861 cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg
     1921 ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga
     1981 catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2041 cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct
     2101 gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa
     2161 gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa
     2221 gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa
     2281 gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa
     2341 gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga
     2401 gggccgccac cacctgttcc tggggcatgg caccggcagc accggcagcg gcagctccgg
     2461 caccgcctcc tccgaggaca acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt
     2521 gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     2581 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
     2641 cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc
     2701 cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     2761 gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg
     2821 cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat
     2881 gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt
     2941 gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga
     3001 gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga
     3061 caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     3121 ctccgagggc cgccaccacc tgttcctgta cggcatggac gagctgtaca agtaataatt
     3181 aattaatgat catacagtgc tctgagtacg ctataatcaa cctctggatt acaaaatttg
     3241 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3301 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3361 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3421 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3481 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3541 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3601 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     3661 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3721 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3781 ctccctttgg gccgcctccc cgcaggatcc actagagcgg ccgcctagag ctcgctatca
     3841 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3901 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3961 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4021 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4081 gcggaaagaa ccagctgggg ctcgactaga gcttgcggaa cccttaagct tctagccaac
     4141 aagctcgtca tcgctttgca gaagagcaga gaggatatgc tcatcgtcta aagaactacc
     4201 cattttatta tatattagtc acgatatcta taacaagaaa atatatatat aataagttat
     4261 cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc ttaaaagtca
     4321 cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca aaatcagtga
     4381 cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg agatgtccta
     4441 aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca agaatgcatg
     4501 cgtcaatttt acgcagacta tctttctagg gttaaaaaag atttgcgctt tactcgacct
     4561 aaactttaaa cacgtcatag aatcttcgtt tgacaaaaac cacattgtgg ccaagctgtg
     4621 tgacgcgacg cgcgctaaag aatggcaaac caagtcgcgc gaggtaccag cttttgttcc
     4681 ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4741 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4801 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4921 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5041 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5521 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5941 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6001 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6061 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6121 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6181 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6241 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6301 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6361 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6421 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6481 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6541 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6601 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6661 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6721 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6781 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6841 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     6901 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6961 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     7021 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     7081 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     7141 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     7201 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     7261 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     7321 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     7381 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     7441 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
     7501 cactataggg cgaattggag ctcggtattc acgacagcag gctgaataat aaaaaaatta
     7561 gaaactatta tttaacccta gaaagataat catattgtga cgtacgttaa agataatcat
     7621 gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt attaatttga
     7681 atagatatta agttttatta tatttacact tacatactaa taataaattc aacaaacaat
     7741 ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct tctataaagt
     7801 aacaaaactt ttaaacattc tctcttttac aaaaataaac ttattttgta ctttaaaaac
     7861 agtcatgttg tattataaaa taagtaatta gcttaaccta tacataatag aaacaaatta
     7921 tacttattag tcagtcagaa acaactttgg cacatatcaa tattatgctc tcgttaatac
     7981 cggtcatggt c
//
