LOCUS       Exported                4694 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Plasmid for making zebrafish apoA-Ia in situ probe.
ACCESSION   .
VERSION     .
KEYWORDS    PCRII apoA-Ia
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  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 4694)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4694
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(923..942)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(924..942)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(949..966)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(949..965)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(958..980)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1106..1534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1193..1212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1403..1424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1878..2672
                     /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(1932..1951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2542..2561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             2690..3550
                     /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(2908..2927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3721..4309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4210..4229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4463..4480
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4597..4618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4633..4663
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4671..4687
                     /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     4676..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 tttccatgta gggctccatg cgagtctgga ggtcctgggt gtgtggggcg
      181 atcttctccc tggcctcctc gacagccttt accagctgct ccttgtactc ctcggtatag
      241 ggggcagcca tggtcctcag gtcctcaaga cgctcggtca gcttagtgcg aacagcctcg
      301 accattggca caactttgga cttggtttcc tcaatgttgg actcgaaggc cttcctaatg
      361 tcgtccatca ggggctccag cttggcacgc agctgctcag cattttggcg gttcagagcg
      421 gagtactcct ggaagacggg ctccagcttc tcacggtact catcgatgtg cttctgcagc
      481 gcggtgtaca gctcggcacg gtggggctcc agcttggatc ggagatcctc aacgtcagtc
      541 atgaccctct cgcgcagctg cttggtgttc tccatcaact gggtggagat ggtctcggcg
      601 tagggggtca aagcctggga ggtggtctgg gcatattcct ggagcttggt caggctctca
      661 gaaagctgga gtttgtactg ctcatagtcg gttccgtcca ggttgtcaag ggctttctca
      721 gcccgatcct tgacctggtt caggtacacc aaggctgctg ccttgtagtg ctccagctgg
      781 gtcggggcat cagcctggaa caaattggcc tgggaaccca aggccaagag aagagtcagt
      841 gcaagagcca cgaaagggcg aattctgcag atatccatca cactggcggc cgctcgagca
      901 tgcatctaga gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
      961 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1021 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1081 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1141 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1201 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1261 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1321 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1381 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1441 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1501 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct
     1561 gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     1621 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
     1681 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
     1741 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     1801 tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag
     1861 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     1921 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     1981 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2041 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2101 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2161 gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca
     2221 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2281 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2341 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2401 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2461 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2521 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2581 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2641 ccttctatcg ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca
     2701 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2761 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2821 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2881 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2941 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3001 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3061 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3121 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3181 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3241 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3301 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3361 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3421 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3481 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3541 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3601 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3661 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3721 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3781 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3841 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3901 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3961 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4021 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4081 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4141 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4201 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4261 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4321 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4381 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4441 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4501 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4561 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     4621 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     4681 ataacaattt caca
//
