LOCUS       sequence-461770-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses zebrafish dgat2 with a C-terminal 3xFLAG tag under control
            of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  AUTHORS   Wilson MH, Hensley MR, Shen MC, Lu HY, Quinlivan VH, Busch-Nentwich 
            EM, Rawls JF, Farber SA
  TITLE     Zebrafish are resilient to the loss of major diacylglycerol 
            acyltransferase enzymes.
  JOURNAL   J Biol Chem. 2024 Nov 6;300(12):107973. doi: 
            10.1016/j.jbc.2024.107973.
  PUBMED    39510175
REFERENCE   2  (bases 1 to 6648)
  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: "J Biol 
            Chem."; date: "2024-11-6"; volume: "300(12):107973. doi"; pages: " 
            10.1016/j.jbc.2024.107973"
COMMENT     Sequence Label: pDEST-CMV-zdgat2FLAG-SV40
FEATURES             Location/Qualifiers
     source          1..6648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(168..192)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(192..210)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(193..210)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(245..1229)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     protein_bind    complement(1253..1273)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(1293..1309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(1682..1844,1845..1856)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     2205..2226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2389..2493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2494..2562,2563..3354)
                     /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(2712..2731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3525..4113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4014..4033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4267..4284
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4401..4422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4437..4442,4443..4460,4461..4467)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4475..4491
                     /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     4480..4502
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4499..4515
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4534..4554
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4536..4554
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     4591..4607
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4591..4607
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(4641..4652,4653..4840)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5279..5413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5429..5445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5453..5473
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    5536..5670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    5672..5696
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5701..5724)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 aaatgtcatg ataaactgca gcacggagat cacctgcaga tgcttcacca ttttggatcg
       61 ggtcagccac gggacggtgg gcaggtcgtg taaggcagag aggatgctgg agcccgagcc
      121 ctttttgaca ccggaatatg cagcaagtat ggtcttcatg tttgcggagc ctgctttttt
      181 gtacaaactt gtctatagtg tcacctaaat caagcttgct ccgacgtccc caggcagaat
      241 ggcggttccc taaacgagca ttgcttatat agacctccca ttaggcacgc ctaccgccca
      301 tttacgtcaa tggaacgccc atttgcgtca ttgcccctcc ccattgacgt caatggggat
      361 gtacttggca gccatcgcgg gccatttacc gccattgacg tcaatgggag tactgccaat
      421 gtaccctggc gtacttccaa tagtaatgta cttgccaagt tactattaat agatattgat
      481 gtactgccaa gtgggccatt taccgtcatt gacgtcaata gggggcgtga gaacggatat
      541 gaatgggcaa tgagccatcc cattgacgtc aatggtgggt ggtcctattg acgtcaatgg
      601 gcattgagcc aggcgggcca tttaccgtaa ttgacgtcaa tgggggaggc gccatatacg
      661 tcaataggac cgcccatatg acgtcaatag gaaagaccat atatagagac cattgacgtc
      721 aatgggggag tggctatggg cggtattagg aagccccata tatggtatat gggaccgccc
      781 attgggaggg gctatctacg tcaataggaa aacccatata tggaatacta tatggcatag
      841 ggccaataca tagtattgaa cctggccaat agccatattg gcatagggcc atattggata
      901 ttgcctatat attgatcctg gcatatagcc aatatggccg ccattattgg caccatgcca
      961 attcaatatg gcggacttgg cacagggcca agacccctcc tcaaacttgg caccgtgcca
     1021 agtagacccc tccccagttg gcacagtgcc aggtccgcca tattgaattg gcatggtgcc
     1081 aagtccgcca tattgaattg gctggggtcc agatccacca tattgaattg gcacgagtcc
     1141 atatacgcca tattgaattg gcacaggtcc agatccacca tattgaattg gcatgagtcc
     1201 atatacgcca tattgaattg gcggtcgact ggcgtaatag cgaagaggcc ttcaactttt
     1261 ctatacaaag ttgatagctt ggcgtaatca tggtcatagc tgtttcctgc tcgagggccc
     1321 atctggcctg tgtttcagac accagggagt ctctgctcac gtttcctgct atttgcagcc
     1381 tctctatcaa gactaataca cctcttcccg catcggctgc ctgtgagagg cttttcagca
     1441 ctgcaggatt gcttttcagc cccaaaagag ctaggcttga cactaacaat tttgagaatc
     1501 agcttctact gaagttaaat ctgaggtttt acaactttga gtagcgtgta ctggcattag
     1561 attgtctgtc ttatagtttg ataattaaat acaaacagtt ctaaagcagg ataaaacctt
     1621 gtatgcattt catttaatgt tttttgagat taaaagctta aacaagaatc tctagttttc
     1681 tttcttgctt ttacttttac ttccttaata ctcaagtaca attttaatgg agtacttttt
     1741 tacttttact caagtaagat tctagccaga tacttttact tttaattgag taaaattttc
     1801 cctaagtact tgtactttca cttgagtaaa atttttgagt actttttaca cctctgtcaa
     1861 gaactcctgg acaaacctct gacctgtgtg gaacagagtg gatatgggtg tctgaacaga
     1921 tattcacgtc ttttgcagat cagagggcat ttctggtgtt gtgcaatgac ctgggtccaa
     1981 caggtaggcg tactcgtatc caagagcatt gttggacagg aggtagtctc cattcctata
     2041 aagaggaagg aatggcacca tgtagtagcc gtcattgtgg ccaattggtg cattggtacg
     2101 tgggtagatg gaccggggag gctgatgagt tcttagccag cgtgggccat cgccctgata
     2161 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2221 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2281 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2341 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     2401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2461 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2521 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2581 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2641 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2701 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2761 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2821 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2881 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2941 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3001 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3061 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3121 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3181 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3241 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3301 gaacgaaata gacagatcgc tgagataggt gcctcactta ttaagcattg gtaactgtca
     3361 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3421 atctaggtga agatcctttt tgataatctc atgaccaaaa tgccttaacg tgagttttcg
     3481 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3541 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3601 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3661 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3721 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3781 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3841 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3901 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3961 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4021 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4081 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4141 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4201 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4261 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4321 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4381 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     4441 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     4501 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     4561 aagctggagc tccaccgcgg tggcggccgc tctagaacta gtggatctgc tgggcttgct
     4621 gaaggtaggg ggtcaagaac cagaggtgta aagtacttga gtaattttac ttgattactg
     4681 tacttaagta ttatttttgg ggatttttac tttacttgag tacaattaaa aatcaatact
     4741 tttactttta cttaattaca tttttttaga aaaaaaagta ctttttactc cttacaattt
     4801 tatttacagt caaaaagtac ttattttttg gagatcactt cattctattt tcccttgcta
     4861 ttaccaaacc aattgaattg cgctgatgcc cagtttaatt taaatgttat ttattctgcc
     4921 tatgaaaatc gttttcacat tatatgaaat tggtcagaca tgttcattgg tcctttggaa
     4981 gtgacgtcat gtcacatcta ttaccacaat gcacagcacc ttgacctgga aattagggaa
     5041 attataacag tcaatcagtg gaagaaaatg gaggaagtat gtgattcatc agcagctgcg
     5101 agcagcacag tccaaaatca gccacaggat caagagcacc cgtggccgta tcttcgcaga
     5161 tctgatctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     5221 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     5281 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5341 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5401 tcatgtctgg atcatcatcg atggtaccgt aaaacgacgg ccagtgaatt atcaactatg
     5461 tataataaag ttggaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5521 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5581 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5641 ctcatcaatg tatcttatca tgtctggatc ccccactttg tacaagaaag ctgggtctca
     5701 tttgtcatca tcgtctttat agtcgtgaat gataagggta tcgctctcat tcagaccgaa
     5761 gcgggtcttg taattgtcaa agagagactt tagggacctg atgtacatgg cgtggtacat
     5821 gtctatgacg tcctgagtcg gctcctctat ctttggcaca gtgataggtt caccaacaac
     5881 tgtagtgata ggtttgcagt aaggcaccag accccaggac tctgggaaaa acagtccaca
     5941 gccatgaaac aaacaagggg caaagcctaa aaacttctga agtttcctct gaattgtacg
     6001 ccaccaagac ccttcttcaa aaatcagctg tttgtaaacc tcattctccc caaaggagta
     6061 aactggcacc agatccgcac cttgctttag agccagtttc acaaagcctt tgcgtttttt
     6121 cagcatgaca gagtttcttc ctggtgcaca gtccagggat tctgcggctc ctccgatcac
     6181 gatgaccaca gcgttgcctg tgccgttact ggacaggagg tagtcgatag agtttcggtt
     6241 tactgggcag attcctccgc acatcaggta ttccctgaac atgggcaggc gaaaatttcc
     6301 agccaatgtg gccagagatg gcttgattcc tgggaacacc ttggtgaatc ctgtggcttc
     6361 tgtcccgaag ttacaaaaag ccccaaaaca caagatgcca tgagggtggt agccaaaaat
     6421 gtagtttcgg ctgggaagca ggttatgcgt cttgatgagt cttatgggaa aatagtctct
     6481 catgtatttc cacacagtcc agtctctcac ccaggtcgat ctgcggcctc cttgtccagg
     6541 ggtgttccag tcatagataa gccaggcaac gtaaagcaca gaaatgaccc agaagtctgt
     6601 gcaaaacatg tacatcagca gcaaagagca agcaatgccc atggtaag
//
