LOCUS       sequence-263497-        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length of nuclear localized tdTomato in pCS expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  AUTHORS   Shin M, Nozaki T, Idrizi F, Isogai S, Ogasawara K, Ishida K, Yuge S,
            Roscoe B, Wolfe SA, Fukuhara S, Mochizuki N, Deguchi T, Lawson ND
  TITLE     Valves Are a Conserved Feature of the Zebrafish Lymphatic System.
  JOURNAL   Dev Cell. 2019 Nov 4;51(3):374-386.e5. doi: 
            10.1016/j.devcel.2019.08.019. Epub 2019 Sep 26.
  PUBMED    31564611
REFERENCE   2  (bases 1 to 5879)
  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"; doi: 
            "10.1016/j.devcel.2019.08.019"; journalName: "Dev Cell"; date: 
            "2019-11-4- 4"; volume: "51"; issue: "3"; pages: "374-386.e5"
COMMENT     Sequence Label: pCS-nlstdTomato
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..1010
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1045..1063
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1045..1062
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1113..1142
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1152..1181
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1191..1220
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1230..1259
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1269..1298
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1332..1361
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1386..1406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1407..2834
                     /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
                     QALKLKDGSHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2844..2900
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    2941..3075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2965..2984)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3019..3038
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3188..3208)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3188..3206)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3227..3243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3227..3243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3240..3262)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3251..3267
                     /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(3275..3281,3282..3299,3300..3305))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3320..3341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3458..3475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3629..4217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3709..3728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4388..5179,5180..5248))
                     /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     5011..5030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5249..5353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5379..5834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5516..5537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5728..5747
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgcgggcctc ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc
       61 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
      121 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
      181 atgccaattc aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac
      241 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
      301 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
      361 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
      421 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
      481 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
      541 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
      601 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
      661 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
      721 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
      781 atctattaat agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc
      841 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
      901 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
      961 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac cgccattctg
     1021 cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc
     1081 tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct gaagaggact
     1141 tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagcaaaagc
     1201 tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga
     1261 atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag agcttgggcg
     1321 acctcaccat ggagcaaaag ctcatttctg aagaggactt gaatttagag atctcgaatt
     1381 ccatgcccaa gaagaagagg aaggtgatgg tgagcaaggg cgaggaggtc atcaaagagt
     1441 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     1501 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1561 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     1621 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     1681 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     1741 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     1801 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     1861 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggca
     1921 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     1981 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     2041 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat ggcaccggca
     2101 gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg gccgtcatca
     2161 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
     2221 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2281 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     2341 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     2401 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     2461 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     2521 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     2581 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     2641 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     2701 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     2761 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg
     2821 acgagctgta caagggatcc ggagccacga acttctctct gttaaagcaa gcaggagacg
     2881 tggaagaaaa ccccggtcct gggctcgagc ctctagaact atagtgagtc gtattacgta
     2941 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3001 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3061 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3121 gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct
     3181 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     3241 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt
     3301 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3361 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3421 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4141 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4201 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4261 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4321 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4381 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4441 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4501 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4561 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4621 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4681 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4741 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4801 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4861 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4921 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4981 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5041 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5101 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5161 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5221 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5281 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5341 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5401 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5461 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5521 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5581 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5641 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5701 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5761 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5821 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcgg
//
