LOCUS       sequence-436965-        4651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  used for TALEN HDR to generate endogenous fusion of HA-Dendra2 to 
            apoBb.1 in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4651)
  AUTHORS   Moll TOC, Kiefer JG, Klemek ML, Wilson MH, Farber SA
  TITLE     Directly Measuring Atherogenic Lipoprotein Kinetics in Zebrafish 
            With the Photoconvertible LipoTimer Reporter.
  JOURNAL   Arterioscler Thromb Vasc Biol. 2025 Aug 21. doi: 
            10.1161/ATVBAHA.125.322969.
  PUBMED    40836916
REFERENCE   2  (bases 1 to 4651)
  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: 
            "Arterioscler Thromb Vasc Biol. 2025 Aug 21. doi: 
            10.1161/ATVBAHA.125.322969."
COMMENT     Sequence Label: apoBb.1-Dendra2
FEATURES             Location/Qualifiers
     source          1..4651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..79)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     179..201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(239..257)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        325..429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             430..1245
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             430..1245
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(503..522)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1416..2004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1905..1924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2158..2175
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2292..2313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2328..2333,2334..2351,2352..2358)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2366..2382
                     /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     2371..2393
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2390..2406
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2390..2406
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(3197..3889)
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     CDS             complement(3902..3928)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4486..4503)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4486..4502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4495..4517)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
       61 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
      121 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      181 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
      241 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
      301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      421 aggaagagta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc
      481 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
      541 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
      601 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
      661 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
      721 accactgcga tccctgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
      781 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
      841 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
      901 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
      961 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     1021 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     1081 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     1141 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     1201 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaactgtc agaccaagtt
     1261 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1321 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1381 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1441 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1501 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1561 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1621 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1681 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1741 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1801 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1861 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1921 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1981 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2041 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2101 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2161 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2221 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2281 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     2341 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     2401 tatgaccatg ctcgagccaa gctcggcgcg ccattgggat ggtaaggcag acatcagttt
     2461 gtaagcaaat ccaaaatatt tttaatgaag attaaatgat tttcagttta tatgtcaaaa
     2521 aggagtagtt aacatgtgtt aacctgaagc tggttaaaat aaggttcaat aaccactaat
     2581 caaagcaatt gatttactag atttttcagg tatgggattg agtaaaatgt tagtagtttt
     2641 ttttacctct acaaaagttc tgataaactt ttttcttcct aatgaaaaaa atagaataaa
     2701 atattttatt tgtaattttt tttaagtcac cagtagtttg cataaaaaac taaaatgtac
     2761 ccatgtacca ctgcttattt ttcattttgt tgcatgttat tacacctgta tattcaatta
     2821 tattatatta ttgaatggtc caaactagtt taaaacactt ctagaatata gaaatgacaa
     2881 gattgggtcg tgttgcataa taaaagttct ttattgatcc ccatccatca tgccattaat
     2941 gttggataca gaacaaggat gacaataggt tgcagttgac atttttgata actagcttag
     3001 tatttcaatc ctcatttata caaatagaat aatagagcag cataacaatg agacatgttt
     3061 tgagtcagta attgctttat atacatgttt gtactatact tcagtgaagt gtccattggc
     3121 ttcgatgtcc aagaatgttc atttcaaaag atggtagtgt ttacttaatt tactttctgt
     3181 tcagaatgac catattttac cacacctggc tgggcagggg gctgtagcgg gccacggcgt
     3241 gctcgtacag cttcaccttg ttgtagtcgc tgtcgttgcc caggatctcg atgcggtggt
     3301 ccacgaagtg ggcgtcgggc agctgcacca ccttcttggc cttgtaggtg gtcttgaagt
     3361 cgcacaggta gtggccgccg ccctccagca gcagggccat gttgatgttg cccaccagca
     3421 ggccgtcgcg cacgtgcagc ttctcggtgc tgggctccca cttcagggtc ttcttctgca
     3481 tcacggggcc gttggggggg aagttggtgc ccttgaagcg cacgttctgg aagaagcagt
     3541 cgccctccag gctgatgtcg ctgcggatgg tgcagatgcc cttgtcctcg aaggtcatgg
     3601 tgcgctccca gctgtagccc tcggggaagc tctgcttgaa gtagtcgggg atgtcctcgg
     3661 ggtacttggt gaacacccgg ttgccgtagt gcacggcggt ggtcaggatg tcgtagctga
     3721 agggcagggg ggcgccctcc ttcacggtca ggttggcggt ctgggtgccc tcgtagggct
     3781 tgcccttgcc ctcgccctcg atcacgaagg cgtggccgtt cacgttgccc tccatgtgca
     3841 ccttcacgcg catgtcctcc ttgatcaggt taattcccgg ggtgttcatt ccaccgccac
     3901 cagcgtaatc tggaacatcg tatgggtact ggaaaaatgg gagaggaagc tcaaattgat
     3961 gtattccatt atttgcattc aacactttcc ctatgttttg agaaatttca tatattttaa
     4021 ttgcagggat tgaaaaacca aagatgactg cctcactaat gtcttctatg atcttcatca
     4081 gctcctggta gaacgtgtta atggccgcgg caatatcttc taataaactt aaattcattt
     4141 tgtccacaaa ttcttgtcct ttttcaacaa cctcttgtaa agtttgtcca agtttcttaa
     4201 gaactacatc aggactttcg atttgcttca ttgcatctgc tatcataact aggttgcttc
     4261 tcaggatact ttgcacattt tgttggaact cagctactgt catgaccttt ccattgggaa
     4321 atgtcaattg aattgtatta aagttatcca tgatgggaga gaagtatacc tctaagttgt
     4381 taccaaaatt cctcagcatt tctacaacaa ctaattggat tttcttgcaa atttcagcca
     4441 gtgtagcttc attcattcca ggcagatccc aatacgcgtc aattcactgg ccgtcgtttt
     4501 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4561 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4621 gcgcagcctg aatggcgaat ggcgcctgat g
//
