LOCUS       sequence-243549-        4190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for making zebrafish apoA-IVb.2 in situ probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4190)
  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 4190)
  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 apoA-IVb.2
FEATURES             Location/Qualifiers
     source          1..4190
                     /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(419..438)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(420..438)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(445..462)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(445..461)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(454..476)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      602..1030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(689..708)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     899..920
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1374..2168
                     /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(1428..1447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2038..2057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2186..2254,2255..3046)
                     /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(2404..2423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3217..3805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3706..3725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3959..3976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4093..4114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4129..4134,4135..4152,4153..4159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4167..4183
                     /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(4172..4190,1..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 gaattcggct tgcagcagtt ttcaaaagtc tgttggtagc atttaatttc tcttggtgta
      181 agattcgtac agagaagcga gctggtcctt caggttctgt gcataggggt ccagcttctc
      241 tttaaggtct tcagcgtagg gtgtaagact ctgctggacc atctgggctc tctctctaat
      301 ctggacctgc agatcctcgg tcaggggagt cacggtcttc tggaactcct aagccgaatt
      361 ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc
      421 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      481 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      541 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      601 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      661 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      721 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      781 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      841 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      901 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      961 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1021 aacgcgaatt ttaacaaaat tcagggcgca agggctgcta aaggaagcgg aacacgtaga
     1081 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     1141 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     1201 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     1261 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     1321 gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     1381 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     1441 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     1501 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     1561 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     1621 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     1681 tgtcatccca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     1741 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     1801 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     1861 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     1921 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     1981 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     2041 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     2101 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     2161 tcttctgaat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2221 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2281 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2341 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2401 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2461 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2521 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2581 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2641 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2701 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2761 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2821 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2881 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2941 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3001 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3061 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3121 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3841 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3901 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3961 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4021 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4081 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     4141 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
//
