LOCUS       sequence-456347-        4820 bp    DNA     circular SYN 15-APR-2025
DEFINITION  zebrafish mogat2 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 4820)
  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 4820)
  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-TOPO-mogat2 in situ probe
FEATURES             Location/Qualifiers
     source          1..4820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             361..1155
                     /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(415..434)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1025..1044
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(1173..1241,1242..2033)
                     /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(1391..1410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2204..2792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2693..2712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2946..2963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3080..3101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3116..3121,3122..3139,3140..3146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3154..3170
                     /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     3159..3181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3178..3194
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3178..3194
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3212..3230
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3212..3229
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4226..4245)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4227..4245)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4252..4269)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4252..4268)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4261..4283)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(4409..4820,1..17)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4496..4515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4706..4727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aaaatttaac gcgaatttta acaaaattca gggcgcaagg gctgctaaag gaagcggaac
       61 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
      121 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
      181 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
      241 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
      301 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
      361 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
      421 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      481 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      541 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      601 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
      661 gatctcctgt catcccacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
      721 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
      781 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
      841 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
      901 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
      961 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     1021 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     1081 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     1141 gacgagttct tctgaattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1201 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1261 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1321 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1381 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1441 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1501 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1561 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1621 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1681 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1741 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1801 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1861 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1921 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1981 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2041 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2101 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2161 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2221 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2281 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2341 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2401 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2461 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2521 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2581 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2641 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2701 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2761 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2821 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2881 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2941 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3001 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3061 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     3121 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     3181 gaaacagcta tgaccatgat tacgccaagc tatttaggtg acactataga atactcaagc
     3241 tatgcatcaa gcttggtacc gagctcggat ccactagtaa cggccgccag tgtgctggaa
     3301 ttcgcccttg gccgaaggtg tactggaaaa ctcctcgcgc atgaaagaga ggaagagaaa
     3361 cacccatgat gcgctgcaga cgctcttgaa tgtatcgcag ccatgtgcca cgaggattct
     3421 gcacctggtc aaacacctcg ttctctccaa acgaatatat aggaaccaaa tctgcaccgt
     3481 gctctattgc aagcttaata aagcctttct tattggccag gtgaacggtg tagtcaccag
     3541 gatgtgcatc caaagcttca ggagctccac cgactgcaat gaccaccgca ttccctccac
     3601 ccttcctccg gagcagataa ctggcactct ccttgtctga tggaaccagc cctgctgaca
     3661 tgatgtaatc tctgaagaag ggggctctga accagagagg cagcatgaga aggttgctct
     3721 ttatcccagg gaagagtttg ctaaatccag tcgcctccgt gcagaaattt gtaaaagcac
     3781 cagccacgag aataccatga ggatggaaac ccatcacata gttcttccta gtgtccaagt
     3841 cagccgtctt tatcagcctg atggggaaat aatctctcat gtactcccac acacgcagtt
     3901 tgcagagata tggcactttt ctccctcctc gtgaaggtgt gtcccagtcg atgtaccacc
     3961 aggtggcata aagcacactg atgagccaga aacgagtgaa gagcaggatg aagaacagga
     4021 caatacagca gggcgcaaga gccaggaatg agaagaccca ttgcagcacg gaagctgtct
     4081 ggagcctccg aaccagaggc acatcagctg gagcaaactc aatcttcatg tctaaaccag
     4141 aagaggacaa gagccgaagg gcgaattctg cagatatcca tcacactggc ggccgctcga
     4201 gcatgcatct agagggccca attcgcccta tagtgagtcg tattacaatt cactggccgt
     4261 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     4321 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     4381 acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg
     4441 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4501 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4561 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4621 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4681 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4741 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4801 aaaaatgagc tgatttaaca
//
