LOCUS       sequence-448284-        5414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  endogenously tag TSC22D4 with N-terminal mNeon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5414)
  AUTHORS   Xiao YX, Lee SY, Aguilera-Uribe M, Samson R, Au A, Khanna Y, Liu Z, 
            Cheng R, Aulakh K, Wei J, Farias AG, Reilly T, Birkadze S, Habsid A,
            Brown KR, Chan K, Mero P, Huang JQ, Billmann M, Rahman M, Myers C, 
            Andrews BJ, Youn JY, Yip CM, Rotin D, Derry WB, Forman-Kay JD, Moses
            AM, Pritisanac I, Gingras AC, Moffat J
  TITLE     The TSC22D, WNK, and NRBP gene families exhibit functional buffering
            and evolved with Metazoa for cell volume regulation.
  JOURNAL   Cell Rep. 2024 Jul 23;43(7):114417. doi: 
            10.1016/j.celrep.2024.114417. Epub 2024 Jul 8.
  PUBMED    38980795
REFERENCE   2  (bases 1 to 5414)
  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: "Cell Rep.";
            date: "2024-07-23"; volume: "43(7):114417. doi"; pages: " 
            10.1016/j.celrep.2024.114417. Epub 2024 Jul 8"
COMMENT     Sequence Label: pYX-N-Puro-mNeon-TSC22D4
FEATURES             Location/Qualifiers
     source          1..5414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             729..1325
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(729..748)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1225..1245
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1335..1391
                     /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             1398..1424
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1398..1424
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1428..1454
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1464..2171
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1719..1728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3193..3209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3193..3209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3206..3228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3217..3233
                     /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(3241..3247,3248..3265,3266..3271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3286..3307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3424..3441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3595..4183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3675..3694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4354..5145,5146..5214))
                     /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     4977..4996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5215..5319)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5387..5405
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cctctctctc tctctctctc
      421 cccctctttt tccagtttgc aaactcagct ctggagttca gcagcaacag cagcaggaaa
      481 aacctgcccc tgctcccccc tcccgccacc tcccctctcc tcttctcccc tcacccagca
      541 ggcacccccg gttcccgcca ggccctcctg ccatgtcgga cccagacgtc cccaggggct
      601 cggatgtccc cgccatgtgg cccccttgtt ccaggggtgc ctgagcccct tcaaggagcc
      661 ccaacccacc cccaaccttg gcccagccct gagccccagg gaccatgggc aagagcaaga
      721 ccggtagcat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
      781 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
      841 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
      901 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
      961 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     1021 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     1081 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     1141 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     1201 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     1261 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     1321 gtgccggatc cggagctact aacttcagcc tgctgaagca ggccggagac gtggaggaga
     1381 accctggacc ttctagatac ccctacgacg tgcccgacta cgccggctat ccgtatgatg
     1441 tcccggacta tgcaggaagc ggaatggtga gcaagggcga ggaggataac atggcctctc
     1501 tcccagcgac acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg
     1561 tgggtcaggg caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca
     1621 agggtgacct ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc
     1681 agtacctgcc ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg
     1741 gataccaagt ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc
     1801 gctacaccta cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc
     1861 ctgctgacgg tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga
     1921 agacttaccc caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa
     1981 atggcaagcg ctaccggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg
     2041 ctaactatct gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca
     2101 agaccgagct caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg
     2161 agctgtacaa gggaggaagc ggaggatctg gcggtaccag cgggggcaag aagaagagta
     2221 gttttcaaat caccagcgtc actacggact atgagggccc tgggagccca ggggcttcgg
     2281 atccccctac cccacagccc ccaaccgggc ccccgccccg cctgcccaat ggggagccca
     2341 gccccgatcc ggggggcaag ggcacccccc ggaatggctc cccaccacct ggggcccctt
     2401 cctcccgttt ccgggtggtg aagctgcccc acggcctggg agagccttat cgccgcggtc
     2461 gctggacgtg tgtggatgtt tatgagcgag acctggagcc ccacagcttc ggcggactcc
     2521 tggagggaat tcgaggggcc tcagggggcg ccgggggcag atctttggat tccaggttgg
     2581 agctggccag cctcggcctg ggcgccccca ccccaccgtc aggcctgtct cagggcccca
     2641 cctcctggct ccgtccaccc cccacctctc ctggacctca ggcccgctcc ttcactgggg
     2701 gactgggcca gctggtggtg cccagcaaag ccaaggcaga gaaaccccca ctgtcggcct
     2761 cctcacccca gcagcgcccc ccagagcctg agaccggtga gagtgcgggc acatcccggg
     2821 ctgccacgcc cctgccctct ctgagggtgg aagcggaggc tgggggctca ggggccagga
     2881 cccctccact gtcccggagg aaagctgtag acatgcggct gcggatggag ttgggtgctc
     2941 cagaagagat ggggcaggta agacctgggt tctagggctg gcccatcagc cctggcctaa
     3001 cctctatctt gagcctcctt cctcctcctc ccccccctct cctcctcctt ctccatcact
     3061 ttataagatc agatagccct cttctctgtg cctgccaggt cactctgaag ccccttcctc
     3121 ctacttcatt ttctgtctca gcctccttcc tgtttttctg caccctctct ctttaagctt
     3181 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3241 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3301 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3421 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4021 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4141 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4201 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4261 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4321 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4381 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4441 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4501 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4561 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4621 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4681 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4741 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4801 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4861 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4921 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4981 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5041 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5101 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5161 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5221 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5281 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5341 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5401 gaggcccttt cgtc
//
