LOCUS       sequence-459801-        4431 bp    DNA     circular SYN 15-APR-2025
DEFINITION  zebrafish mogat3b 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 4431)
  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 4431)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-mogat3b in situ probe
FEATURES             Location/Qualifiers
     source          1..4431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     188..207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     441..458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    575..596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(611..616,617..634,635..641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    649..665
                     /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     654..676
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     673..689
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     673..689
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        707..725
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     707..724
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1332..1351)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1333..1351)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1358..1375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1358..1374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1367..1389)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1515..1943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1602..1621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1812..1833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2287..3081
                     /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(2341..2360)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2951..2970
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3099..3167,3168..3959)
                     /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(3317..3336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4130..4431,1..287)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
       61 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      301 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      361 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      421 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      481 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      541 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      601 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      661 aacaatttca cacaggaaac agctatgacc atgattacgc caagctattt aggtgacact
      721 atagaatact caagctatgc atcaagcttg gtaccgagct cggatccact agtaacggcc
      781 gccagtgtgc tggaattcgc ccttacggtc cactcttgca atcttagccg gacttttccg
      841 ccttcctctc ttcagggaat acattttagc tgcaggattg ctgcctgtca gtaaggcaag
      901 tttggattat ttgctgagcc aaactggtgt gggtaacgct gtggtcatca tcattggtgg
      961 agcagaggaa tcactgacct cctctacagg agtaaacact gtggtcataa aacacaggaa
     1021 aggctttgta cgactggctc ttgaacacgg ggcagacctg gttccagtgt actcttttgg
     1081 agagaacgag ctgttcccgc aggtggttct gtctgagggc agtgtaggtc gacggctgca
     1141 ggttcttttt aagcagatta tgggttttgc tccatgcatc ttcacaggag gccggtggct
     1201 tcttctgccg tacaaacttc cagtcaccac tgtggtgggc tgccccataa acgtcctctg
     1261 gtaagggcga attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag
     1321 ggcccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     1381 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1441 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1501 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1561 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1621 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1681 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     1741 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     1801 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     1861 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     1921 ttaacaaaaa tttaacgcga attttaacaa aattcagggc gcaagggctg ctaaaggaag
     1981 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     2041 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     2101 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     2161 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     2221 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg
     2281 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2341 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2401 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2461 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2521 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2581 gggcaggatc tcctgtcatc ccaccttgct cctgccgaga aagtatccat catggctgat
     2641 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2701 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2761 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     2821 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2881 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2941 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3001 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3061 cttcttgacg agttcttctg aattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3121 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3181 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3241 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3301 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3361 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3421 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3481 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3541 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3601 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3661 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3721 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3781 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3841 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3901 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3961 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4021 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4081 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4141 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4201 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4261 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4321 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4381 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag g
//
