LOCUS       sequence-115854-        7498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of tdTomato from the broadly active CAG 
            promoter/enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7498)
  AUTHORS   Miura H, Inoko H, Tanaka M, Nakaoka H, Kimura M, Gurumurthy CB, Sato
            M, Ohtsuka M
  TITLE     Assessment of Artificial MiRNA Architectures for Higher Knockdown 
            Efficiencies without the Undesired Effects in Mice.
  JOURNAL   PLoS One. 2015 Aug 18;10(8):e0135919. doi: 
            10.1371/journal.pone.0135919. eCollection 2015.
  PUBMED    26285215
REFERENCE   2  (bases 1 to 7498)
  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: "PLoS One.";
            date: "2015-08-18"; volume: "10(8):e0135919. doi"; pages: " 
            10.1371/journal.pone.0135919. eCollection 2015"
COMMENT     Sequence Label: pART (pT-2)
FEATURES             Location/Qualifiers
     source          1..7498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        320..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        625..900
                     /label=chicken beta-actin promoter
     intron          901..1909
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1833..1855
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1977..1986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1983..3413
                     /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"
     intron          3447..4019
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4151..4170)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4216..4271
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4270..4289)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4627..4644)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4627..4643)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4636..4658)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4856..5311
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4943..4962)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5153..5174
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5328..5347)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5447..5469
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5507..5525)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5593..5697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5698..5766,5767..6558)
                     /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(5916..5935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6729..7317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7218..7237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7471..7488
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtcgactct agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      661 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      721 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      781 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      841 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      901 ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg ccgcccgccc
      961 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     1021 ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc
     1081 cttaaagggc tccgggaggg ccctttgtgc gggggggagc ggctcggggg gtgcgtgcgt
     1141 gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc ccggcggctg tgagcgctgc
     1201 gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga ggggagcgcg gccgggggcg
     1261 gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     1321 gggggggtga gcagggggtg tgggcgcggc ggtcgggctg taaccccccc ctgcaccccc
     1381 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtgcg gggcgtggcg
     1441 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     1501 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg ccccggagcg ccggcggctg
     1561 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     1621 acttcctttg tcccaaatct ggcggagccg aaatctggga ggcgccgccg caccccctct
     1681 agcgggcgcg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     1741 gtgcgtcgcc gcgccgccgt ccccttctcc atctccagcc tcggggctgc cgcaggggga
     1801 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1861 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1921 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgggatcca ccggtcgcca
     1981 ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg
     2041 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     2101 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     2161 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
     2221 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     2281 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     2341 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     2401 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     2461 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     2521 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
     2581 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     2641 gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc agctccggca
     2701 ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc ttcaaggtgc
     2761 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2821 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     2881 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg
     2941 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     3001 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca
     3061 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc
     3121 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     3181 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     3241 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca
     3301 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
     3361 ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag taaagtctag
     3421 attcgaagga tcctgagaac ttcagggtga gtttggggac ccttgattgt tctttctttt
     3481 tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg ttgtttagaa
     3541 tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc tttcactttc
     3601 tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta actttttcgt
     3661 taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat ttgtcagatt
     3721 gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta tattgtactt
     3781 cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata tttctgcata
     3841 taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacatc ctggtcatca
     3901 tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg ataaaatact
     3961 ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     4021 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttcactcctc
     4081 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     4141 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     4201 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     4261 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     4321 atttggttta gagtttggca acatatgcca tatgctggct gccatgaaca aaggtggcta
     4381 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     4441 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     4501 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     4561 ccagtcatag ctgtccctct tctcttatgg agatccctcg aggctagcgg taccgagctc
     4621 gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4681 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4741 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     4801 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4861 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4921 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4981 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5041 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     5101 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5161 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     5221 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5281 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     5341 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5401 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5461 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5521 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5581 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5641 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5701 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5761 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5821 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5881 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5941 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6001 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6061 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6121 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6181 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6241 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6301 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6361 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6421 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6481 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6541 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6601 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6661 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6721 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6781 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6841 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6901 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6961 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7021 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7081 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7141 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7201 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7261 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7321 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7381 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7441 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
