LOCUS       sequence-243550-        4819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for making zebrafish apoEb in situ probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4819)
  AUTHORS   Otis JP, Zeituni EM, Thierer JH, Anderson JL, Brown AC, Boehm ED, 
            Cerchione DM, Ceasrine AM, Avraham-Davidi I, Tempelhof H, Yaniv K, 
            Farber SA
  TITLE     Zebrafish as a model for apolipoprotein biology: comprehensive 
            expression analysis and a role for ApoA-IV in regulating food 
            intake.
  JOURNAL   Dis Model Mech. 2015 Mar;8(3):295-309. doi: 10.1242/dmm.018754. Epub
            2015 Jan 29.
  PUBMED    25633982
REFERENCE   2  (bases 1 to 4819)
  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"; doi: 
            "10.1242/dmm.018754"; journalName: "Dis Model Mech"; date: 
            "2015-03"; volume: "8"; issue: "3"; pages: "295-309"
COMMENT     Sequence Label: PCRII apoEb
FEATURES             Location/Qualifiers
     source          1..4819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1048..1067)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1049..1067)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1074..1091)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1074..1090)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1083..1105)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1231..1659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1318..1337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1528..1549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2003..2797
                     /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(2057..2076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2667..2686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2815..2883,2884..3675)
                     /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(3033..3052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3846..4434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4335..4354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4588..4605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4722..4743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4758..4763,4764..4781,4782..4788)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4796..4812
                     /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     join(4801..4819,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat agaatactca
       61 agctatgcat caagcttggt accgagctcg gatccactag taacggccgc cagtgtgctg
      121 gaattcgccc tttgtccagt aaaggacagt gctagtccaa ttgagtccag ttgttaacca
      181 ctttatcaac ctctcacaga atgaaaaaag cacatttcag taacacaaag cctcccttct
      241 tctttctgtt tggtgttaac actaggtaag tgtggagctc ttaagcctga gtgggaagag
      301 ctgctgtgtc ctccttgatc ttgtccatga cttcctgcag ctgctcgcgg atcttctggg
      361 agtagggggc gaggaggctg gtcagctcat ccacacggcc ctccagagag gtgcgtaggc
      421 tctcggctgt ctcctccagc ttctccttca gagctccagc ctggacctcc aactgctcgc
      481 tcacctcctg cgccagggcc ttcatcagct cgctaatggc gcccagtttc tgggtggcgc
      541 catcctgggc ctggctcatg tatggctgga aacggtcctt cactgcatca gcattttggg
      601 aagcgcggga ctgcatctca ctcatgtagg ttgctacggt gttgcggatc tcctctgtgt
      661 ccttgttcag gcgtttcttc agtttgcgtg tgtaggtgcc gacacggttc ttcacgtcat
      721 ctgcattctg ctccatcatg gtcttcagct cttgcaggta ctgagtgctg cgttccttag
      781 cgtcggtcat gtcagtttgg agttttccag ccaggagctg aagatcttta ctgagctgac
      841 cagcagcatc agaggcgtat ggggtcatct gggtttggag attttcactg tatgagctca
      901 gttcagccat ggtgtcagta attagtgtgt caagctctct gctgagctgg gagcccttga
      961 tgttttgcac catgccgtaa gggcgaattc tgcagatatc catcacactg gcggccgctc
     1021 gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     1081 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1141 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1201 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     1261 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1321 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1381 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1441 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1501 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1561 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1621 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaatt cagggcgcaa
     1681 gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg
     1741 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
     1801 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
     1861 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     1921 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
     1981 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2041 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2101 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2161 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     2221 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2281 attgggcgaa gtgccggggc aggatctcct gtcatcccac cttgctcctg ccgagaaagt
     2341 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2401 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     2461 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2521 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2581 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2641 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     2701 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     2761 catcgccttc tatcgccttc ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagt
     2821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3001 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3661 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3721 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3781 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4501 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4561 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4621 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4681 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4741 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4801 agcggataac aatttcaca
//
