LOCUS       sequence-252464-        6724 bp    DNA     circular SYN 14-APR-2025
DEFINITION  RNA isolation from machrophages via TRAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6724)
  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 6724)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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(mpeg1:EGFP-Mmu.Rpl10a)
FEATURES             Location/Qualifiers
     source          1..6724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2045..2054
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2051..2053,2054..2056,2057..2767)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2051..2053,2054..2056,2057..2767)
                     /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(2096..2117)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2357..2376)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2704..2725
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3568..3689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3605..3624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3659..3678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3980..4000)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3980..3998)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4019..4035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4019..4035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4032..4054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4043..4059
                     /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(4067..4073,4074..4091,4092..4097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4112..4133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4250..4267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4421..5009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4501..4520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5180..5971,5972..6040))
                     /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     5803..5822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6041..6145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6308..6329)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta cttagggaaa
       61 attttactca attaaaagta aaagtatctg gctagaatct tacttgagta aaagtaaaaa
      121 agtactccat taaaattgta cttgagtatt aaggaagtaa aaggctagca agatctgctc
      181 gagtgttgga gcacatctga catctgaaat tatttgaaag cagcagaaat gtttttacag
      241 ctattacaaa atttacttaa caagaatcct ttaaaatatg acaatttatt gccaataatt
      301 tacagtaagt tttgcagtac tttaatatta caaattatac aactgtgctg gcacacaaaa
      361 aataacttaa tccagcacaa caacaacaac aacaactaca aaaagcatat ttccttttgc
      421 tttttcttgt tcactcaact tttggaaaca tcaactgcac tgacctaaaa ttatacagta
      481 tgtcagtaat acaaaaacac ttagttaggt taacataaga ttattagttt tctggagagg
      541 tgaactactc tgtaagaaac ctttttaagt tgtgccaact aaacattttt atcagcagaa
      601 ctacaatgct tgatgagtga acaaaaaaac aaacaaacct tatttccttt tgttttttct
      661 tgtgtactta acattttttt cacctttatt tgaaacgaca gtagggagta tggacaggaa
      721 agtgttggga gaggagaggg atgagcatag aaccaggagg taggaatcaa actcaggtca
      781 ctatgaacac cggagtgcca ggtgtcaaca cactaaccac tacaccatca actccaacat
      841 ttacttgaaa acatcagttg agtgaacaaa aaactcttgt ggaaattggt actgagaaat
      901 aatacattat cacaacaaac aaaacagaga caaatgttaa caaaacaatt attagaaaca
      961 gaattagaaa ccggacagga ggtaaaacaa taatgataaa caagacaagg atcaaaacat
     1021 ggcaggacaa agacagtttt ttgttcactc aacctcagat attttaggtc agtgcaactg
     1081 atgtttacaa aggttgtatg catgtcaaca ccaatggtgt agtgattagt gtgttgacac
     1141 aaggcactcc agtgttcaca ctgatgctgg tttgattccc acatcgatgt cctttgctaa
     1201 tcctgcctct ctccccatgc tttcctgtct atactctcta ctgttgtgtc aattaaaggt
     1261 gaaaaaagtt aagcacaaaa gaaaaatcta aggatataag gattatgttt ttttttgttc
     1321 actcaaaatc aggcattcca gttctgcaga caaaaaatgt tcagttggca caacttaaaa
     1381 agttttgtta cagagtagtt cacctctcca gaaaactaat aatcttatgt taacccaact
     1441 aaatgttttt actgaaaaat aattgaatgt cagtgcagct gatgtttttt taaaagtgga
     1501 gagaacaaaa aaagtgaaag gaaataagga ttcattgttg taccgactta tttttttgaa
     1561 gaaaattctg ctatagggct ttcatgatcc cctatgctca cagcagtttt aggagatgct
     1621 tataataaag aggatttgaa aaacctgcta atcttgtaca tttctaaaac aagaagccaa
     1681 ccaatttcaa ttcctcctct ttcaaagcca atggaaaatt taaaacgcac agtctcttgc
     1741 gtcatcaaaa ccattctgaa gactcatcaa aatccttcgt agctttaaat ataaatctat
     1801 tattgaacag atcatacttc acaaaaaata taaatatatt tgattcattc acttcatgaa
     1861 attgtccttc aacttaacgt ttgaacgcaa ggtaccaaaa cacaaagagg aacctcatag
     1921 tctcatgacc cttcccccac ggatttgaaa ctgagaagtg gttttattga ggaactcagc
     1981 aaaatctata atcaccatat ttgattgaaa atcacattag tgcaggagac agcaaaagga
     2041 tcccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2101 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2161 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2221 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2281 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2341 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2401 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2461 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2521 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2581 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2641 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2701 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2761 gtacaagtcc ggccggactc agatctcgag ctcaagcttc gaattcagca gcaaagtctc
     2821 acgcgacacc ctgtacgagg cggtgcggga agtcctgcac gggaaccagc gcaagcgccg
     2881 caagtttctg gagacggtgg agctgcagat cagcctgaag aactacgacc ctcagaagga
     2941 caaacgtttc tcgggcaccg tcaggctcaa gtccacccca cgccccaagt tctcggtgtg
     3001 cgttctgggg gaccagcagc actgtgatga agccaaggcc gtggatatcc cccacatgga
     3061 catcgaggcg ctcaagaagc ttaacaaaaa caagaagttg gtcaagaagc tggctaagaa
     3121 gtacgatgcc tttttggcct ctgagtctct gattaagcag atcccacgta tcctgggccc
     3181 aggcctaaac aaggctggca agttcccctc cctgctgaca cacaatgaaa acatggtggc
     3241 caaagtggat gaggtgaaat cgacaatcaa gttccagatg aagaaggtgc tgtgtttggc
     3301 cgtcgctgtt ggccacgtga agatgaccga tgatgagcta gtctacaaca ttcatctggc
     3361 tgtcaatttc ttggtgtcct tgcttaagaa aaactggcaa aacgtgcggg ctctgtacat
     3421 caagagcacc atgggcaagc cccagcgtct gtattagtct agatcataat cagccatacc
     3481 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3541 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3601 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3661 ggtttgtcca aactcatcaa tgtatcttaa ggcgagatcc tgcggccgca agggaaaata
     3721 gaatgaagtg atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta
     3781 aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa
     3841 ttgtactcaa gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa
     3901 attactcaag tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga
     3961 tccactagat cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4021 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4081 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4141 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4201 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4261 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4321 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5161 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5341 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5401 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5461 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5581 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5701 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5821 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5881 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6061 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6121 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     6181 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     6241 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     6301 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     6361 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     6421 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     6481 agcttgacgg ggaaagccgg ccacaatgac ggctactaca tggtgccatt ccttcctctt
     6541 tataggaatg gagactacct cctgtccaac aatgctcttg gatacgagta cgcctacctg
     6601 ttggacccag gtcattgcac aacaccagaa atgccctctg atctgcaaaa gacgtgaata
     6661 tctgttcaga cacccatatc cactctgttc cacacaggtc agaggtttgt ccaggagttc
     6721 ttga
//
