LOCUS       sequence-243569-        4741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for making zebrafish apoEa in situ probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4741)
  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 4741)
  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 apoEa
FEATURES             Location/Qualifiers
     source          1..4741
                     /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(970..989)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(971..989)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(996..1013)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(996..1012)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1005..1027)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1153..1581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1240..1259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1450..1471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1925..2719
                     /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(1979..1998)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2589..2608
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2737..2805,2806..3597)
                     /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(2955..2974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3768..4356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4257..4276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4510..4527
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4644..4665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4680..4685,4686..4703,4704..4710)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4718..4734
                     /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(4723..4741,1..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 ttgagctcct gcattcgtga gtccagtctc tcctttattt cctcagcctg
      181 gcttctcagc gcttctctca gattcaccag cttctgttga gcgcgctcgt gcatcgacgt
      241 gaagtaaggc tccagacgat ccttcacatc ttccacgctc tgtggagcac gagactgaac
      301 ctcctcaagg tatgttgaca tttttttgtt gatctcctgg atgtccttgt taaatcgttt
      361 ccttaatttc ttggcgtagg agttagcttt gtttttgagc tcatcagagt tctgctccac
      421 catggtccgg agctcctctg tgtattcagt gacacgatcc tttgcctcct ccatgtgtgt
      481 gcggagcttg gtgaccaaca actgcagatc ttcgttgaac ttctgggctc tctcctgcgc
      541 gcaaggggcc agtttggagt gcagatcatc tctgtacatt tgcagctcca tcatgctgtc
      601 agagatcagt gtgtctaatt ctctgccgag ctgagtggcc ttgatgttgt ccctcatttc
      661 ctcagctcta gcggtcagtt cagaaacatg gttccagaac tgatccacgg cctcttccca
      721 gcggcttctt ggctcatcct gaaacaggaa tcgcccctgg cagcctgaaa tgacggcgag
      781 cgcaacaatt acagccacaa acttcatgat tgtatttgga cagtacacca gacagatggg
      841 ctctgtaagg aagtcctttg gaaagagctg ttatttatac tgcttggatc gcagtcgttc
      901 aagggcgaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
      961 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     1021 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     1081 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1141 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1201 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1261 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1321 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1381 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1441 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     1501 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1561 aacaaaaatt taacgcgaat tttaacaaaa ttcagggcgc aagggctgct aaaggaagcg
     1621 gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     1681 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     1741 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     1801 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     1861 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
     1921 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1981 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2041 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2101 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2161 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2221 gcaggatctc ctgtcatccc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     2281 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     2341 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     2401 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     2461 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     2521 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2581 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     2641 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2701 tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2761 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2821 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2881 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2941 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3001 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3061 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3121 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3181 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3241 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3301 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3361 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3421 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3481 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3541 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3601 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3661 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3721 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3781 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3841 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3901 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3961 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4021 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4081 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4141 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4201 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4261 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4321 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4381 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4441 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4501 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4561 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4621 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4681 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4741 a
//
