LOCUS       sequence-461745-        6967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses zebrafish dgat1a with a C-terminal FLAG tag under control 
            of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6967)
  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 6967)
  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: pcDNA3.1+ dgat1a-FLAG
FEATURES             Location/Qualifiers
     source          1..6967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2426..2449
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2558..2575)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2564..2788
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2834..3262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2921..2940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3131..3152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3271..3291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3276..3605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3456..3591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3518..3537
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3672..4466
                     /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(3726..3745)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4336..4355
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4640..4761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4677..4696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4731..4750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4810..4826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4810..4826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4823..4845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4834..4850
                     /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(4858..4864,4865..4882,4883..4888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4903..4924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5041..5058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5212..5800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5292..5311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5971..6762,6763..6831))
                     /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     6594..6613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6832..6936)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgcaaacat gggcgacaga aacgagaagg gctcggcgaa
      961 gcaccggcgg aggactacga tctccggaga ggcggcggtg actcaagcca aacgcggcgg
     1021 cgcagcagag acgctgagcg ggcaggtgaa ggagaaagag ccgcagaaac acgagaatgc
     1081 agccggtaga cagaagaacc acagcgatgc tggagaggac acattcagct gtcacaagct
     1141 gcaggagtca ctgctcagtt cggccagtgg ttttagcaac tacagaggaa tcctcaactg
     1201 gtgtgtggtc atgctggttt tgtctaacgc acggctcgtg ctggagaacc ttataaaata
     1261 tggcgttttg gtggacccca tccagatcat ctctctgttc gtgaaggacc cctgcagctg
     1321 gccggctctg tgcctgatca tcgtcactaa cgtcttcata atggcggctc tgtacaccga
     1381 gagaaagctg tctgtgggca ccatctctga gagaaccgga tctttccttc actgcatcaa
     1441 tctgtctgca ctgttgtttg tgccggcagg aacggtgcta agcctgagct ccgtcacacc
     1501 agtgggcggt gtgatggcac tgagtatctg cactgttctc ttcctcaagc tttactcgta
     1561 cacagatgtt aataagtggt gccgagaaca gagacaagcc aaagctcgaa ccctgtctcg
     1621 ctctcactcc tgtccgtccg tgcacaaagc aaatggcaca gcgggataca cacacgtgac
     1681 gtatccggga aacctcacac acagagacat ttattatttt gcttttgcac cgactctgtg
     1741 ttacgagctg aattttccgc gctcaccacg aatacgcaaa cgtttcctac tgcggcgact
     1801 gctggagatg ctgtttctaa tgcaactgat ggtcggatta atacagcagt ggatggttcc
     1861 gacgattcag aactctatga aaccctttca ggacatggat ttctccagaa tggtggagcg
     1921 gatgctgaaa ctcgctgtgc ccaatcactt tatctggctc atctttttct attggtattt
     1981 ccactcctct atgaacttcg tagcagagat catgcagttc ggggaccgcg agttttaccg
     2041 cgactggtgg aactctgaaa cgattcctta tttctggtca aactggaaca ttcctgttca
     2101 caagtggtgc ctacgacact tttataagcc catgttgatg cgaggagcaa atcgattagc
     2161 agctcagatc gccgtcttct tcctctccgc ttttttccac gagtatttag tgagtgttcc
     2221 tctcaagatg tttcggctct gggcgtttct gggtatgatg ctgcaggtcc cgctggccat
     2281 tttggttggc cgatacctga ggggtaacta tggcaacgcc gccgtgtgga tgtctctgat
     2341 catcggccag ccaatcgcag tgctcatgta cgtccacgac tactacgtgt tgcattatgg
     2401 cagcgcgtct gaggccgcag cgccggacta taaagacgat gatgacaaat gaggtaccga
     2461 gctcggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg
     2521 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2581 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2641 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2701 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2761 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2821 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2881 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3181 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3241 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3481 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3541 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3601 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3661 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3721 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3781 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3841 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3901 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3961 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4021 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4081 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4141 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4201 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4261 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4321 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4381 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4441 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4501 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4561 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4621 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4741 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4801 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4861 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4921 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4981 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5641 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6961 tgacgtc
//
