LOCUS       sequence-456350-        3663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway middle entry vector containing zebrafish dgat2 with a 
            C-terminal 3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3663)
  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 3663)
  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: pME-zdgat2-CDS-FLAG
FEATURES             Location/Qualifiers
     source          1..3663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(9..95)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     144..166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     158..175
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     159..175
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    191..290
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1382..1405
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(1410..1509)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1526..1545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1527..1545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1550..1566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1550..1566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1668..1687)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1679..2488
                     /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(1746..1765)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2635..3223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3124..3143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3377..3394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3553..3580)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
       61 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
      121 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
      181 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
      241 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcaaacat
      301 gaagaccata cttgctgcat attccggtgt caaaaagggc tcgggctcca gcatcctctc
      361 tgccttacac gacctgccca ccgtcccgtg gctgacccga tccaaaatgg tgaagcatct
      421 gcaggtgatc tccgtgctgc agtttatcat gacatttctt accatgggca ttgcttgctc
      481 tttgctgctg atgtacatgt tttgcacaga cttctgggtc atttctgtgc tttacgttgc
      541 ctggcttatc tatgactgga acacccctgg acaaggaggc cgcagatcga cctgggtgag
      601 agactggact gtgtggaaat acatgagaga ctattttccc ataagactca tcaagacgca
      661 taacctgctt cccagccgaa actacatttt tggctaccac cctcatggca tcttgtgttt
      721 tggggctttt tgtaacttcg ggacagaagc cacaggattc accaaggtgt tcccaggaat
      781 caagccatct ctggccacat tggctggaaa ttttcgcctg cccatgttca gggaatacct
      841 gatgtgcgga ggaatctgcc cagtaaaccg aaactctatc gactacctcc tgtccagtaa
      901 cggcacaggc aacgctgtgg tcatcgtgat cggaggagcc gcagaatccc tggactgtgc
      961 accaggaaga aactctgtca tgctgaaaaa acgcaaaggc tttgtgaaac tggctctaaa
     1021 gcaaggtgcg gatctggtgc cagtttactc ctttggggag aatgaggttt acaaacagct
     1081 gatttttgaa gaagggtctt ggtggcgtac aattcagagg aaacttcaga agtttttagg
     1141 ctttgcccct tgtttgtttc atggctgtgg actgtttttc ccagagtcct ggggtctggt
     1201 gccttactgc aaacctatca ctacagttgt tggtgaacct atcactgtgc caaagataga
     1261 ggagccgact caggacgtca tagacatgta ccacgccatg tacatcaggt ccctaaagtc
     1321 tctctttgac aattacaaga cccgcttcgg tctgaatgag agcgataccc ttatcattca
     1381 cgactataaa gacgatgatg acaaatgaga cccagctttc ttgtacaaag ttggcattat
     1441 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1501 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     1561 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     1621 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     1681 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     1741 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     1801 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     1861 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     1921 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     1981 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2041 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2101 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2161 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2221 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2281 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2341 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2401 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2461 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     2521 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     2581 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2761 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3241 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3301 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3361 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3421 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     3481 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt
     3541 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
     3601 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
     3661 gct
//
