LOCUS       sequence-429345-        4203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenously tag human TMEM115 with a C-terminal 3xHA tag in 
            addition to cytosolic mNG for cell sorting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4203)
  AUTHORS   Hollingsworth LR, Veeraraghavan P, Paulo JA, Harper JW
  TITLE     Spatiotemporal proteomic profiling of cellular responses to NLRP3 
            agonists
  JOURNAL   bioRxiv 2024.04.19.590338
REFERENCE   2  (bases 1 to 4203)
  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: "bioRxiv 
            2024.04.19.590338"
COMMENT     Sequence Label: hTMEM115-TEV-3xHA-P2A-mNeonGreen HDR template
FEATURES             Location/Qualifiers
     source          1..4203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(52..108)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(118..144)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(148..174)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(178..204)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(205..225)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(923..940)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(923..939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(932..954)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1053..1157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1158..1226,1227..2018)
                     /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(1376..1395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    2118..2142
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      2259..2847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2748..2767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3000..3022
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3019..3035
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3019..3035
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(join(3547..4203,1..51))
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      3993..4002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcatgtgtc gctgggagag aggccatgtt atcctcctcg cccttgctca ccggtccagg
       61 attctcttcg acatctccgg cttgtttcag cagagagaag tttgttgcgc cggatcctgc
      121 gtagtctggg acgtcatagg ggtacccggc atagtcgggc acgtcgtagg gatagccagc
      181 gtaatccgga acatcatacg ggtagccctg gaagtacagg ttttccagcg tcgggggagc
      241 tgcctcgaag gtgattagac tggattctgg ggcagccccc ttccctgggg gtgtgggagc
      301 tttgtctgag ggcagggggc tgtccacctt ggccccagac tcctcttcat catcatccat
      361 gctgggccag atggactggt cttccactct cttcagccgc tcattgaggg ctttgagtgc
      421 cagttgcctg gggccagagg agagaggaaa ggtcactctg ggtttggcct tggctcatgc
      481 cctctaccct tccgcagcag gctgactcct caccgtacca cctagactcc tcccccagaa
      541 tcctatgttt ccccttcacc atgcagagcc tgggactccc cagagattgg tttaggaccc
      601 actctgccat cagaatgata acccaaaggc tctaagaatc ttccctccat agctgaggca
      661 cctccgggat gggaaggtca gctgggtgtc tgccagactg cccacgaggc cagccaccag
      721 gaatggttct gagggtggta tctaagggtc agtggaacta gtgccctagg gcagatgggt
      781 ggaacggcaa gttctggagt ttccaggagt ttctgcctgg gtgctggcat cacatccaag
      841 ggaagggtta ttaattctcc ccctctcctg aggtcaggcg gaatggcact tccttattcc
      901 agatgcgtgc ggattatgca tgactggccg tcgttttaca acgtcgtgac tgggaaaacc
      961 ctggcgcttg tggtggatca cagttattgg agacttcctt ggccgtacga tgggttttgt
     1021 ttcaggtatc tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1081 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1141 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1201 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1261 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1321 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1381 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1441 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1501 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1561 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1621 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1681 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1741 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1801 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1861 ccggtgagcg tggctctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1921 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1981 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2041 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2101 ttgaattgcc attatcaaca agtttgtaca aaaaagcagg ctccgaattg gtatcacgag
     2161 gacgatttga agccccctac ccatttacgc tgacttgacg ggacggcggc tttgttgaat
     2221 agagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2401 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2881 gccttttgct ggccttttgc tcacatgttc tttccagcga catatgccat acgccgaact
     2941 gaactccacc taatgtcttg gagattgtcg gatagcgcgg aataggattg tgccctctga
     3001 gcggataaca atttcacaca ggaaacagct atgaccatga cacatgcatc caccatcgca
     3061 gacttatcat cactgagcct ccacctagcc tcctgacagg aacagtcttt ctggggatgg
     3121 gcagggatgt gcagccccag gtcggctcct gcatttgctg ggcccccaga acatttttgg
     3181 caaatccttg ccttggctca gggctctcgg caaccagttt cagatgtgag ggccggccca
     3241 ggagctgtgg catcagtgtc cctgcagcca cttgcttggg gctgccagct gatgtacatt
     3301 cttacctcag atgagggcct ggactggccg atgcctcaga ggctccgggc ctggcatagt
     3361 ggttgtctgg agttggaacc aggtagcaag agtcctgggt agtgttggcg agaaacagca
     3421 gaaggccatg gaaagcccca gcaggccttc ttccctctcc tcagagcagt caggtgccct
     3481 ggagtagctg agaggatgtc aaggggggct tgggagggga ggtgccacac tcaaggtggt
     3541 ctggagttac ttgtacagct cgtccatgcc catcacatcg gtaaaggcct tttgccactc
     3601 cttgaagttg agctcggtct tggagtgctt gagctccgtc ttacggaaca cgtacatcgg
     3661 ctggttcttc agatagttag ccgccattgg cttggcaaag gtgtaggtgg tccgcgcagt
     3721 gctccggtag cgcttgccat ttccagtggt gtaactccac ttaaaggtac tgatgatggt
     3781 tttgtcgttg gggtaagtct tcttcgacct gcaccagtcc gcagcggtca gcgagttggt
     3841 catcacagga ccgtcagcag ggaaaccagt ccccttcacc tgggcctctc ctttgatgtg
     3901 gcttccctcg taggtgtagc ggtagttaac agtaagggag gcaccatctt caaactgcat
     3961 tgtgcgatgg acttggtatc cggagccatc taccatggcg gcctggaaag gcgacatccc
     4021 gtcagggtag ggcaggtact gatggaagcc atacccgata tgagggacca gaatccaggg
     4081 ggagaactgg aggtcaccct tggtggactt caggtttaac tcctcataac catcatttgg
     4141 attgccggtg ccctgaccca ccatgtcaaa gtccacaccg ttgatggagc caaagatgtg
     4201 taa
//
