LOCUS       sequence-252467-        7345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RNA isolation from neutrophils via TRAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7345)
  AUTHORS   Lam PY, Harvie EA, Huttenlocher A
  TITLE     Heat shock modulates neutrophil motility in zebrafish.
  JOURNAL   PLoS One. 2013 Dec 19;8(12):e84436. doi: 
            10.1371/journal.pone.0084436. eCollection 2013.
  PUBMED    24367659
REFERENCE   2  (bases 1 to 7345)
  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: "PLoS One.";
            date: "2013-12-19"; volume: "8(12):e84436. doi"; pages: " 
            10.1371/journal.pone.0084436. eCollection 2013"
COMMENT     Sequence Label: Tg(lyz:EGFP-Mmu.Rpl10a)
FEATURES             Location/Qualifiers
     source          1..7345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1614..1623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1620..1622,1623..1625,1626..2336)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1620..1622,1623..1625,1626..2336)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1926..1945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2273..2294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3137..3258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3174..3193)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3228..3247
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(3382..3569,3570..3581)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3636..3656)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3636..3654)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3675..3691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3675..3691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3688..3710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3699..3715
                     /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)."
     promoter        complement(join(3723..3729,3730..3747,3748..3753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3768..3789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3906..3923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4077..4665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4157..4176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4836..5627,5628..5696))
                     /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     5459..5478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5697..5801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5964..5985)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6381..6392,6393..6530)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 gcacactggg ttgcattaag ctgcagtttt ttcacaacag ttttccacat tgtaacagtc
       61 agtctccaaa ttattccatc agtcctcaaa tttagtcatt tgaggccatg aaaacactta
      121 aaactacatg aaggtagttt ggcattaggt tttagacttg ttcttgtttg gcatattgca
      181 acctgtgtaa aaatgatttg atttttattt tttttttttt tttttcagaa agaaacggtc
      241 caaaaagagt tatgtgaaca tttatccatg tgtttaatgt tttttaaatt aaaaaaaaac
      301 atttttatta aaccacttgt gactaaacct tttttagtta ttctattttg tgacgagtct
      361 gaatggccaa agtactcttg aatccaaaat ctttatagta aaatgtttaa tcttacttgc
      421 agtgttgggt ataatttgta acaaagttga attccatatc tgctgagtga aaaagtaaag
      481 taagagattg ctgtaaatat ttaggttatt tatttattaa tacaatatag aaagtaacaa
      541 aacacaatgt aaaaaatcaa taaagaaagt aaatattaaa agtaatgata aactaatgat
      601 gtaaacaata ttagcaacaa agtttgtaca attcagggag ggtaaagaat ttcacctaat
      661 agtcaaactc tgtattaaat aatataaaat ctttgttctt agtctgaaga ggagactgtg
      721 agagaaagct tactggttgt taaattagaa aatttgtaac ttgcagatta aaatgattta
      781 aataatttta taaaggccat gtttgaaaca aatgtaaaaa cataataaat gttaagaatt
      841 tcagatattg agattttaca atttaaaaaa agcaataaag taatagcaga gacctacact
      901 cttaaagtaa attacccaac tctgcttact agtaactttt ggaatagtta ctagtatcca
      961 ttgaacaaat actagaatct aattaaattg tgtgagtgtc caatgaaaag attcttgtat
     1021 ttaaaacagt accagtatct atatatgtac aaaatttagt gctagcaaaa tgcaaataaa
     1081 tactactgca tataatggat tgcattgtaa atcgactcgc cataatacat ttatttccta
     1141 tttttacttt taacaaaaaa taagttcact gagtatttca tgtcattaaa tcgcattaaa
     1201 cctcttgcat gctttatgta cactagtgca gttcgctaaa tgaccaccgg tgtcactaat
     1261 aacacgatcg tttctgaatt ctgcattaaa cgccatttat acgttgctct tattagtaaa
     1321 caatttctcc atcatgagct cgaccagtat tgcatatttc atcctcttaa atccagagtt
     1381 gtttctttta atcactttct cttcatgata aaagcagttt ttggttttca gaagtttaca
     1441 acaaacccag tcagttattg cctcagcaaa cactggagca actgcacact tgtgaaaagc
     1501 ttcttgcgaa acacaaacct ttctctggac ttataaataa aggcattcag cagctccacc
     1561 atacacctcc actggtcttg ataaagcaga tatcagcagt gatacaggta cccgccacca
     1621 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1681 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1741 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1801 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1861 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1921 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1981 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2041 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2101 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2161 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2221 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2281 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     2341 gccggactca gatctcgagc tcaagcttcg aattcagcag caaagtctca cgcgacaccc
     2401 tgtacgaggc ggtgcgggaa gtcctgcacg ggaaccagcg caagcgccgc aagtttctgg
     2461 agacggtgga gctgcagatc agcctgaaga actacgaccc tcagaaggac aaacgtttct
     2521 cgggcaccgt caggctcaag tccaccccac gccccaagtt ctcggtgtgc gttctggggg
     2581 accagcagca ctgtgatgaa gccaaggccg tggatatccc ccacatggac atcgaggcgc
     2641 tcaagaagct taacaaaaac aagaagttgg tcaagaagct ggctaagaag tacgatgcct
     2701 ttttggcctc tgagtctctg attaagcaga tcccacgtat cctgggccca ggcctaaaca
     2761 aggctggcaa gttcccctcc ctgctgacac acaatgaaaa catggtggcc aaagtggatg
     2821 aggtgaaatc gacaatcaag ttccagatga agaaggtgct gtgtttggcc gtcgctgttg
     2881 gccacgtgaa gatgaccgat gatgagctag tctacaacat tcatctggct gtcaatttct
     2941 tggtgtcctt gcttaagaaa aactggcaaa acgtgcgggc tctgtacatc aagagcacca
     3001 tgggcaagcc ccagcgtctg tattagtcta gatcataatc agccatacca catttgtaga
     3061 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3121 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3241 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3301 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc ggatcctgcg
     3361 gccgcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt gactgtaaat
     3421 aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa
     3481 aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata atacttaagt
     3541 acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac cccctacctt
     3601 cagcaagccc agcagatcca ctagatcttt tgttcccttt agtgagggtt aattgcgcgc
     3661 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5821 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5881 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5941 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     6001 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6061 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6121 ggagcccccg atttagagct tgacggggaa agccggccac aatgacggct actacatggt
     6181 gccattcctt cctctttata ggaatggaga ctacctcctg tccaacaatg ctcttggata
     6241 cgagtacgcc tacctgttgg acccaggtca ttgcacaaca ccagaaatgc cctctgatct
     6301 gcaaaagacg tgaatatctg ttcagacacc catatccact ctgttccaca caggtcagag
     6361 gtttgtccag gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa
     6421 agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct
     6481 tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa
     6541 aaggctagca agatctgctc gagatacatt tgattagaag agtttgactt gggttgatta
     6601 tagcaaatag tggaacacaa acagattttg aaatattctt gcaaatttat ttattaaatc
     6661 cattcaaata ttgtcatttt ttgtactgtg taatttatat tgatttgctc atacaatatt
     6721 tacatttcgc ttaaacaata cactattatt tatatttaaa aatagctata tataaatcaa
     6781 atgtatagct cttttttaca caattaatca aagctaaaat atttgtcaca caaatacagt
     6841 aagccacttc acaatgatat cttcaaaatc tgtgcttgac tgaatcattt catttattgt
     6901 tttgtaattc acattatacc ttacgttctc caggcacagt ttgaattgtt gataaatgaa
     6961 gataatacat ctgatttaat tagtctcctt tgtttataat aactttaatc taatttgata
     7021 gtaataattc cagcaataca tccattacaa taaaaatcgc ttttttattt tgcacattag
     7081 tttgccggta tttgtttaaa gtttcaccat acttttaaat catgtgctat ttgaaaaggg
     7141 ctgttatgac tggaattaaa tgtttttttt tatatcagtg ttttagattg aagtacaata
     7201 tttataggta aaattcacca gcatatgctt aaagatagcc atgctccagg tccttgtgtc
     7261 cttctaaggt ctgtttgtgg tttaagctgt ttgttgacac acgcttgtct tctccatttt
     7321 tctgtgaagt tattgtcagt ttatt
//
