LOCUS       sequence-456342-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zebrafish dgat2 anti-sense in situ probe, linearize with BamHI, T7 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  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 4767)
  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: pCRII-dgat2 in situ probe
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..51)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     242..261
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     474..496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     488..505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     489..505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     512..531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        512..530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1474..1492)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1475..1492)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1510..1526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1510..1526)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1523..1545)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1534..1550
                     /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(1558..1564,1565..1582,1583..1588))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1603..1624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1741..1758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1912..2500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1992..2011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2671..3462,3463..3531))
                     /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     3294..3313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3549..4343)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3660..3679)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4270..4289
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(join(4687..4767,1..348))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
       61 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
      121 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
      181 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
      241 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
      301 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
      361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
      421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      481 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg
      541 ccctctagat gcatgctcga gcggccgcca gtgtgatgga tatctgcaga attcgccctt
      601 gccaccaaga cccttcttca aaaatcagct gtttgtaaac ctcattctcc ccaaaggagt
      661 aaactggcac cagatccgca ccttgcttta gagccagttt cacaaagcct ttgcgttttt
      721 tcagcatgac agagtttctt cctggtgcac agtccaggga ttctgcggct cctccgatca
      781 cgatgaccac agcgttgcct gtgccgttac tggacaggag gtagtcgata gagtttcggt
      841 ttactgggca gattcctccg cacatcaggt attccctgaa catgggcagg cgaaaatttc
      901 cagccaatgt ggccagagat ggcttgattc ctgggaacac cttggtgaat cctgtggctt
      961 ctgtcccgaa gttacaaaaa gccccaaaac acaagatgcc atgagggtgg tagccaaaaa
     1021 tgtagtttcg gctgggaggc aggttatgcg tcttgatgag tcttatggga gaatagtctc
     1081 tcatgtattt ccacacagtc cagtctctca cccaggtcga tctgcggcct ccttgtccag
     1141 gggtgttcca gtcatagata agccaggcaa cgtaaagcac agaaatgacc cagaagtctg
     1201 tgcaaaacat gtacatcagc agcaaagagc aagcaatgcc catggtaaga aatgtcatga
     1261 taaactgcag cacggagatc acctgcagat gcttcaccat tttggatcgg gtcagccacg
     1321 ggacggtggg caggtcgtgt aaggcagaga ggatgctgga gcccgagccc tttttgacac
     1381 cggaatatgc agcaaagggc gaattccagc acactggcgg ccgttactag tggatccgag
     1441 ctcggtacca agcttgatgc atagcttgag tattctatag tgtcacctaa atagcttggc
     1501 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1561 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1621 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1681 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1741 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1801 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1861 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1921 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1981 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2041 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2101 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2161 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2221 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2281 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2341 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2401 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2461 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2521 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2581 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2641 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2701 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2761 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2821 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2881 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2941 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3001 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3061 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3181 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3541 ttttcaattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     3601 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3661 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     3721 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3781 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
     3841 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     3901 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     3961 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     4021 gcaaggtggg atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     4081 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     4141 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
     4201 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     4261 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     4321 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt
     4381 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     4441 ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa
     4501 accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg
     4561 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt
     4621 ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg ccctgaattt
     4681 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4741 tcggcaaaat cccttataaa tcaaaag
//
