LOCUS       sequence-343156-        5343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PLIN2-tdTOMATO fusion (zebrafish lipid droplet reporter) in middle 
            entry vector for Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5343)
  AUTHORS   Lumaquin D, Johns E, Montal E, Weiss JM, Ola D, Abuhashem A, White 
            RM
  TITLE     An in vivo reporter for tracking lipid droplet dynamics in 
            transparent zebrafish.
  JOURNAL   Elife. 2021 Jun 11;10. pii: 64744. doi: 10.7554/eLife.64744.
  PUBMED    34114952
REFERENCE   2  (bases 1 to 5343)
  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: "Elife."; 
            date: "2021-06-11"; volume: "10. pii: 64744. doi"; pages: " 
            10.7554/eLife.64744"
COMMENT     Sequence Label: pME-PLIN2-tdTOMATO
FEATURES             Location/Qualifiers
     source          1..5343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1453..2883
                     /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
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(2900..2999)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3016..3035)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3017..3035)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3040..3056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3040..3056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3158..3177)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3169..3978
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3236..3255)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4125..4713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4614..4633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4867..4884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5043..5070)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5162..5248)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5297..5319
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5311..5328
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5312..5328
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc
      121 atgagctttc ttctgtactt gaaactgcac aacagatttt cggctaacat gggttctatg
      181 gaggatgtaa ataaccagaa tgtggttgcc cgaattgttc accttcccct ggttgtttcc
      241 acctatgaca tgatgtcgaa cgtgtacgtc aacaccaaga acaaccaccc atacctgaag
      301 tctgtgtgcg aggttgctga gaatggtgtc aaatccataa cctcagctgc ctttcacagc
      361 gctttaccaa tcattggcaa actggagcct cagatctcta tggcaaatga cctggcgtgc
      421 aaaggacttg ataagataga aaagactctg ccgatcctac acaagccatc tgaacagatt
      481 atggccaatg ctaaagatgc agtgacagga gccaaggagg ctgtcagcgg gactgtgagc
      541 ggtgccaagg agtctgtctc catcacccta aatgatgtga tggaccgaac acgcggggca
      601 gtgcagggca gcgtggagaa gaccaagact gtggtcagcg gtggagtcca gacagtgatg
      661 gacagcagga tgctcaaact ggtgtccagc agagtggaca atgcactgag cacatctgag
      721 acgctgctgg agcactactt gcctgaaaat gaggaggaaa aagaaggtga agagaggaat
      781 actgaaggat tggaaaatga cagcgatttg gccaattatt atgtccgact gggctctctg
      841 tccattaaag tcagagacag gacttatcag agagccgtgg agaaagttcg tcatgccgtg
      901 cagcgcggac aggagtccat ttcacagctt aaccacacca tggatataat cgaatttacc
      961 aggaagaata ttgatgggtc taaccagaag ctgcaggaga aactgggttc actaatgggc
     1021 tggaagagcg tttctcaaag ccaagaaaca gagagcgaaa cagacagtga agctgagcaa
     1081 attgagtcgc gcacactgac catcgcccac agcctcagcc agcagcttca gagcacatgt
     1141 gtggtgctgg tgtccagtct caggggtctc cctcagcacg tccagctcca ggtgggctct
     1201 gtcagccgct ccgcattgga gatctacagc agcttcagta aagcggccgt gctgggggat
     1261 ctgtccagca ccgtcctgtc ctccagccgc tcacagctca gcagaatcac agagtccatg
     1321 gacaacgtga tggactacct gctcaacaac actcctctca actggctggt cggaccgttt
     1381 tacccacgtg tggagcgcgc cgttgaaaac gaggccaaat gtagcaaaca gaggaccctt
     1441 caagtcactg aaatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
     1501 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1561 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     1621 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     1681 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     1741 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     1801 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     1861 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     1921 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     1981 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     2041 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     2101 cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc
     2161 agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc
     2221 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     2281 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     2341 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     2401 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     2461 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     2521 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     2581 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     2641 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccgctac
     2701 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     2761 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2821 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     2881 taaaagggtg ggcgcgccga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     2941 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     3001 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     3061 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     3121 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     3181 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     3241 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     3301 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     3361 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     3421 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     3481 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     3541 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     3601 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     3661 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     3721 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     3781 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     3841 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     3901 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     3961 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     4021 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     4081 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4141 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4201 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4261 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4321 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4381 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4441 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4501 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4561 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4621 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4681 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4741 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4801 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4861 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4921 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4981 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     5041 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     5101 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     5161 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     5221 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     5281 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     5341 ccc
//
