LOCUS       sequence-296716-        5287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse Kir3.2 fused to mCherry and Streptag II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5287)
  AUTHORS   Qiao P, Liu Y, Zhang T, Benavides A, Laganowsky A
  TITLE     Insight into the selectivity of Kir3.2 toward 
            phosphatidylinositides.
  JOURNAL   Biochemistry. 2020 May 6. doi: 10.1021/acs.biochem.0c00163.
  PUBMED    32372643
REFERENCE   2  (bases 1 to 5287)
  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: 
            "Biochemistry. 2020 May 6. doi: 10.1021/acs.biochem.0c00163."
COMMENT     Sequence Label: pPIC-mouseKir3.2-mC-S
FEATURES             Location/Qualifiers
     source          1..5287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1537..1557
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2284..2307
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      2387..2633
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2648..3060
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3067..3114
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3133..3507
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3573..3821
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3589..3607)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    3905..3952
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     rep_origin      complement(4674..5262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4754..4773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttaaacgc tgtcttggaa cctaatatga caaaagcgtg atctcatcca agatgaacta
       61 agtttggttc gttgaaatgc taacggccag ttggtcaaaa agaaacttcc aaaagtcggc
      121 ataccgtttg tcttgtttgg tattgattga cgaatgctca aaaataatct cattaatgct
      181 tagcgcagtc tctctatcgc ttctgaaccc cggtgcacct gtgccgaaac gcaaatgggg
      241 aaacacccgc tttttggatg attatgcatt gtctccacat tgtatgcttc caagattctg
      301 gtgggaatac tgctgatagc ctaacgttca tgatcaaaat ttaactgttc taacccctac
      361 ttgacagcaa tatataaaca gaaggaagct gccctgtctt aaaccttttt ttttatcatc
      421 attattagct tactttcata attgcgactg gttccaattg acaagctttt gattttaacg
      481 acttttaacg acaacttgag aagatcaaaa aacaactaat tattcgaaac gaaaatggca
      541 aaacgtaaga tccaacgtta cgttcgtaag gacggtaagt gcaatgtgca tcatggtaat
      601 gtaagagaaa cctacagata cctaacggac attttcacca ctttggtgga tttgaaatgg
      661 cgtttcaact tattgatatt tgttatggta tacaccgtta cgtggctatt tttcggtatg
      721 atttggtggc taattgcata tataagaggt gacatggatc acatcgaaga cccctcctgg
      781 actccatgtg ttaccaatct aaacggcttt gtttccgcat tcttgttttc catcgaaact
      841 gagaccacaa ttggttacgg ttaccgtgta attacagata aatgcccaga gggtatcatc
      901 ttgttactaa ttcaatccgt tttaggctcc attgttaatg ctttcatggt cggttgcatg
      961 tttgtgaaaa tttctcagcc caaaaagagg gctgaaactc ttgttttctc cacacatgca
     1021 gttatatcaa tgagggatgg caagttatgc ctgatgtttc gtgtcggaga cctaagaaat
     1081 tcacatatag tggaagcaag tattagagct aaactgataa aaagtaagca aacatccgag
     1141 ggcgagttca tcccattgaa tcagtctgac attaacgtgg gatactatac tggcgacgac
     1201 cgtctttttt tagttagtcc acttatcatt tcccacgaaa ttaatcagca gtcccctttt
     1261 tgggagattt ccaaggctca attgcctaag gaagaactag agattgtcgt gattcttgaa
     1321 ggcatcgtcg aagccactgg catgacgtgc caagcacgtt cctcttacat cacatccgag
     1381 atactatggg gatatagatt cacgcccgtc ttaaccatgg aagatggatt ctacgaagta
     1441 gattataaca gtttccatga gacctatgag acgagtactc cttccctatc tgcaaaggaa
     1501 ttggctgaac tggctaatag agccgagggt cctggcgaaa acctgtattt tcagggtact
     1561 agtgtctcta agggagaaga ggacaatatg gctattatca aggagttcat gagatttaaa
     1621 gtccatatgg agggctctgt taatggtcat gaatttgaaa ttgaaggtga aggagaagga
     1681 agaccttatg agggaaccca aacggctaag ctgaaagtaa ctaaaggcgg cccacttcct
     1741 ttcgcatggg atatactgtc tccacaattc atgtatggct ctaaagctta tgtaaagcac
     1801 cccgccgaca ttcccgatta tctaaagtta tcctttcccg agggatttaa atgggaaagg
     1861 gttatgaact tcgaggatgg cggcgtggta actgttacgc aggactcctc ccttcaagat
     1921 ggtgaattta tttacaaagt gaagcttcgt ggtactaatt tcccaagtga cggtcctgtg
     1981 atgcagaaga agactatggg ttgggaagct agttccgaga ggatgtaccc cgaagatggt
     2041 gccctgaagg gtgagattaa gcaacgtcta aaattaaaag acggcggtca ctatgacgct
     2101 gaagtaaaga ccacatataa ggcaaaaaag cccgtccagc ttcctggcgc ctacaatgtt
     2161 aatatcaagt tggacatcac ctcccataac gaagattaca ctatcgtcga gcaatacgag
     2221 agggcagagg gaagacattc cacaggtggt atggacgagc tatataaact cgagggcgct
     2281 tcctggtcac atccacagtt tgagaaatga gtttgtagcc ttagacatga ctgttcctca
     2341 gttcaagttg ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca
     2401 gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta
     2461 tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag
     2521 cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga
     2581 tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt
     2641 ttgtgcggat ctcccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc
     2701 agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact
     2761 aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa
     2821 agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat
     2881 cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat gacctcccat
     2941 tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt
     3001 ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag
     3061 ggcggtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3121 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     3181 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     3241 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     3301 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     3361 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     3421 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     3481 cacttcgtgg ccgaggagca ggactgacac gtccgacgcg gcccgacggg tccgaggcct
     3541 cggagatccg tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
     3601 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     3661 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     3721 aatttttctt tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     3781 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctggag accaacatgt
     3841 gagcaaaagg ccagggcgac gtaaacggcc acaagtttat ggccgtgatg acctgtgtct
     3901 tcgtggtttg tctggtcaac caccgcggtc tcagtggtgt acggtacaaa cccaaagcag
     3961 cacgacacgg caactacaag acccgcgccg agggcatgtt ccccaaagcg ataccacttg
     4021 aagcagtggt actgcttgtg ggtacactct gcgggtgtga agttcgaggg cgacaccctg
     4081 gtgaaccgca tcgagctgaa gggcatcttc caactcgctt aattgcgagt ttttatttcg
     4141 tttatttcaa ttaaggtaac taaaaaactc cttttacaca tgaagcagca cgacttcttc
     4201 aagcgttaag ggattttggt catgagatca gatctaacat ccaaagacga aaggttgaat
     4261 gaaacctttt tgccatccga catccacagg tccattctca cacataagtg ccaaacgcaa
     4321 caggagggga tacactagca gcagaccgtt gcaaacgcag gacctccact cctcttctcc
     4381 tcaacaccca cttttgccat cgaaaaacca gcccagttat tgggcttgat tggagctcgc
     4441 tcattccaat tccttctatt aggctactaa caccatgact ttattagcct gtctatcctg
     4501 gcccccctgg cgaggttcat gtttgtttat ttccgaatgc aacaagctcc gcattacacc
     4561 cgaacatcac tccagatgag ggctttctga gtgtggggtc aaatagtttc atgttcccca
     4621 aatggcccaa aactgacagt ttaaacccgt aaaaaggccg cgttgctggc gtttttccat
     4681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5101 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5221 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5281 tctacgg
//
