LOCUS       sequence-456346-        4728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zebrafish dgat1b 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 4728)
  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 4728)
  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-dgat1b in situ probe
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     193..212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     425..447
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     439..456
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     440..456
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     463..482
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        463..481
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1386..1404)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1387..1404)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1422..1438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1422..1438)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1435..1457)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1446..1462
                     /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(1470..1476,1477..1494,1495..1500))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1515..1536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1653..1670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1824..2412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1904..1923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2583..3374,3375..3443))
                     /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     3206..3225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3461..4255)
                     /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(3572..3591)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4182..4201
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(join(4599..4728,1..299))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(join(4709..4728,1..2))
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
       61 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
      121 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
      181 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
      241 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
      301 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      361 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      421 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
      481 ggcgaattgg gccctctaga tgcatgctcg agcggccgcc agtgtgatgg atatctgcag
      541 aattcgccct tgaaaatgcg accgctacga cagaatgagt tgccacaagt tgcaggagtc
      601 gatgttgagt tcagcgagca gcttcaaaaa ctacagaggg atcctgaact ggtgtgtggt
      661 aatgctggtt ttgagcaacg cacgtctctt tttagaaaac cttctcaggt atggcatttt
      721 agtggacccc attcaggtgg tgtctttgtt tctcaaggac ccttatagct ggcctgctgc
      781 atgtctggtt attgtttgta atgcttttat acttgtggca ctgtacaccg agagaaagct
      841 agccatgggg tcgatcagtg aaaaggtcgg cctgctgatc tacatcttta atttaacaat
      901 catcttgtgt tttccaatgg ttgttgtttt aaaactacca tctattacgc cagttggagg
      961 ggcatttgcc ttaggaatct acaccatcct gattttgaaa ctgtattcat acaaagacgt
     1021 taacaagtgg tgcagagaga gaacacaagc gaaggcccgc agcctctcta gatctctttc
     1081 gtgtccatca cctccatcta catccagcag tatgcagtct tatgtgtctt acccaggaaa
     1141 cctctctctc agggatatat actattttgt ttttgctccg accctgtgct acgagcttaa
     1201 tttcccccga tctgaatcaa tccgaatggg atttctcctc aggaggctct ttgagatgtt
     1261 gcttctcact cagttattgg ttggtttaac gcagcagtgg atggtgaagg gcgaattcca
     1321 gcacactggc ggccgttact agtggatccg agctcggtac caagcttgat gcatagcttg
     1381 agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1441 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1501 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1561 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3421 acacggaaat gttgaatact catactcttc ctttttcaat tcagaagaac tcgtcaagaa
     3481 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     3541 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     3601 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     3661 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     3721 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     3781 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     3841 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     3901 catcagccat gatggatact ttctcggcag gagcaaggtg ggatgacagg agatcctgcc
     3961 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     4021 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     4081 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     4141 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     4201 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     4261 acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg
     4321 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     4381 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     4441 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     4501 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc
     4561 gcttccttta gcagcccttg cgccctgaat tttgttaaaa ttcgcgttaa atttttgtta
     4621 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4681 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtcc
//
