LOCUS       sequence-456345-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zebrafish dgat1a anti-sense in situ probe, linearize with NotI, SP6 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  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 4758)
  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-TOPO-dgat1a in situ probe
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(514..1305,1306..1374))
                     /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     1137..1156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(1392..2186)
                     /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(1503..1522)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2113..2132
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(2530..2958)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2640..2661)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2852..2871
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3084..3106
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3098..3115
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3099..3115
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3122..3141
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3122..3140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4075..4093)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4076..4093)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4111..4127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4111..4127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4124..4146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4135..4151
                     /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(4159..4165,4166..4183,4184..4189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4204..4225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4342..4359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4513..4758,1..343))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4593..4612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
       61 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      481 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      541 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      601 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      661 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      721 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      781 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      841 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      901 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      961 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1021 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1081 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1141 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1201 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1261 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1321 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1381 cctttttcaa ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     1441 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     1501 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     1561 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     1621 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     1681 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     1741 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     1801 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     1861 ggagcaaggt gggatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     1921 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     1981 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     2041 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     2101 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     2161 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     2221 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     2281 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat
     2341 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     2401 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     2461 cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccctgaa
     2521 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2581 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2641 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2701 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2761 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2821 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2881 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2941 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3001 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     3061 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     3121 gtaatacgac tcactatagg gcgaattggg ccctctagat gcatgctcga gcggccgcca
     3181 gtgtgatgga tatctgcaga attcgccctt cggccagtgg ttttagcaac tacagaggaa
     3241 tcctcaactg gtgtgtggtc atgctggttt tgtctaacgc acggctggtg ctggagaacc
     3301 ttataaaata tggcgttttg gtggacccca tccagatcat ctctctgttc gtgaaggacc
     3361 cctgcagctg gccggctctg tgcctgatca tcgtcactaa cgtcttcata atggcggctc
     3421 tgtacaccga gagaaagctg tctgtgggca ccatctctga gagaaccgga tctttccttc
     3481 actgcatcaa tctgtctgca ctgttgtttg tgccggcagg aacggtgcta agcctgagct
     3541 ccgtcacacc agtgggcggt gtgatggcac tgagtatctg cactgttctc ttcctcaagc
     3601 tttactcgta cacagatgtt aataagtggt gccgagaaca gagacaagcc aaagctcgaa
     3661 ccctgtctcg ctctcactcc tgtccgtccg tgcacaaagc aaatggcaca gcgggataca
     3721 cacacgtgac gtatccggga aacctcacac acagagacat ttattatttt gctttcgcac
     3781 cgactctgtg ttacgagctg aattttccac gctctccgcg aatacgcaaa cgtttcctac
     3841 tgcggcgact gctggagatg ctgtttctaa tgcagctgat ggtcggatta atacagcagt
     3901 ggatggttcc gacgattcag aactctatga aaccctttca ggacatggat ttctccagaa
     3961 tggtggagcg gatgctgaaa ctcgctgtgc ccaataaggg cgaattccag cacactggcg
     4021 gccgttacta gtggatccga gctcggtacc aagcttgatg catagcttga gtattctata
     4081 gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4141 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4201 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4261 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4321 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4381 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4441 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4501 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4561 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4621 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4681 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4741 ggtcgttcgc tccaagct
//
