LOCUS       sequence-337988-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of full-length, human GIRK4 with C-terminal mCherry 
            StrepTag II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4731)
  AUTHORS   Qiao P, Schrecke S, Lyu J, Zhu Y, Zhang T, Benavides A, Laganowsky A
  TITLE     Insight into the Phospholipid-Binding Preferences of Kir3.4.
  JOURNAL   Biochemistry. 2021 Dec 21;60(50):3813-3821. doi: 
            10.1021/acs.biochem.1c00615. Epub 2021 Nov 30.
  PUBMED    34846128
REFERENCE   2  (bases 1 to 4731)
  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"; doi: 
            "10.1021/acs.biochem.1c00615"; journalName: "Biochemistry"; date: 
            "2021-12-21- 21"; volume: "60"; issue: "50"; pages: "3813-3821"
COMMENT     Sequence Label: pACE-GIRK2-mC-S
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        108..199
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     124..141
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      204..213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1488..1508
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2235..2258
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     polyA_signal    2285..2419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2322..2341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2376..2395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2540..2705
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2895..3483
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3384..3403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  3618..3841
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3908..4441)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4630..4635,4636..4652,4653..4658))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(join(4729..4731,1..31))
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 acttcgtata gcatacatta tacgaagtta tctgtaacta taacggtcct aaggtagcga
       61 gtttaaacac tagtatcgat tcgcgaccta ctccggaata ttaatagatc atggagataa
      121 ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt
      181 aataaaaaaa cctataaata accgccacca tggccaagct gacagaatcc atgactaacg
      241 tcctggaggg cgactccatg gatcaggacg tcgaaagccc agtggccatt caccagccaa
      301 agttgcctaa gcaggccagg gatgacctgc caagacacat cagccgagat cggaccaaaa
      361 ggaaaatcca gaggtacgtg aggaaagacg gaaagtgcaa tgttcatcac ggcaacgtga
      421 gggagaccta tcgctacctg accgatatct tcaccacatt agtggacctg aagtggagat
      481 tcaacctatt gatttttgtc atggtttaca cagtgacctg gctctttttt ggaatgatct
      541 ggtggttgat cgcatacata cggggagaca tggaccacat agaggacccc tcctggactc
      601 cttgtgttac caacctcaac gggttcgtct ctgctttttt attctcaata gagacagaaa
      661 ccaccattgg ttatggctac cgggtcatca cagataaatg cccggaggga attattcttc
      721 tcttaatcca atctgtgttg gggtccattg tcaatgcatt catggtggga tgcatgtttg
      781 taaaaatctc tcaacccaag aagagggcag agaccctggt cttttccacc catgcagtga
      841 tctccatgcg ggatgggaaa ctgtgcctga tgttccgggt aggggacctt aggaattccc
      901 acattgtgga ggcttccatc agagccaagt tgatcaaatc caaacagacc tcggaggggg
      961 agttcatccc gttgaaccag acggatatca acgtagggta ttacacgggg gatgaccgtc
     1021 tgtttctggt gtcaccgctg atcattagcc atgaaattaa ccaacagagt cctttctggg
     1081 agatctccaa agcccagctg cccaaagagg aactggaaat tgtggtcatc ctagaaggaa
     1141 tggtggaagc cacagggatg acatgccaag ctcgaagctc ctacatcacc agtgagatcc
     1201 tgtggggtta ccggttcaca cctgtcctga ccctggagga cgggttctac gaagttgact
     1261 acaacagctt ccatgagacc tatgagacca gcaccccatc ccttagtgcc aaagagctgg
     1321 ccgagttagc cagcagggca gagctgcccc tgagttggtc tgtatccagc aaactcaacc
     1381 aacatgcaga actggagact gaagaggaag aaaagaacct cgaagagcaa acagaaagaa
     1441 atggtgatgt ggcaaacctg gagaatgaat ccaaagttgg taccagcgaa aacctgtatt
     1501 ttcagggtac tagtgtctct aagggagaag aggacaatat ggctattatc aaggagttca
     1561 tgagatttaa agtccatatg gagggctctg ttaatggtca tgaatttgaa attgaaggtg
     1621 aaggagaagg aagaccttat gagggaaccc aaacggctaa gctgaaagta actaaaggcg
     1681 gcccacttcc tttcgcatgg gatatactgt ctccacaatt catgtatggc tctaaagctt
     1741 atgtaaagca ccccgccgac attcccgatt atctaaagtt atcctttccc gagggattta
     1801 aatgggaaag ggttatgaac ttcgaggatg gcggcgtggt aactgttacg caggactcct
     1861 cccttcaaga tggtgaattt atttacaaag tgaagcttcg tggtactaat ttcccaagtg
     1921 acggtcctgt gatgcagaag aagactatgg gttgggaagc tagttccgag aggatgtacc
     1981 ccgaagatgg tgccctgaag ggtgagatta agcaacgtct aaaattaaaa gacggcggtc
     2041 actatgacgc tgaagtaaag accacatata aggcaaaaaa gcccgtccag cttcctggcg
     2101 cctacaatgt taatatcaag ttggacatca cctcccataa cgaagattac actatcgtcg
     2161 agcaatacga gagggcagag ggaagacatt ccacaggtgg tatggacgag ctatataaac
     2221 tcgagggcgc ttcctggtca catccacagt ttgagaaata ataatgaatg caattgttgt
     2281 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2401 atcttatcat gtctggatct gatcactgct tgagcctaga agatccggct gctaacaaag
     2461 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactatca taacccctag
     2521 ggtataccca tctaattgga accagataag tgaaatctag ttccaaacta ttttgtcatt
     2581 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     2641 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     2701 ccacaaccgg ttgacttggg tcaactgtca gaccaagttt actcatatat actttagatt
     2761 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2821 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2881 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2941 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3001 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3061 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3121 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3181 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3241 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3301 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3361 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3421 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3481 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3541 atgttctttc ctgcgttatc ccctgattga cttgggtcgc tcttcctgtg gatgcgcaga
     3601 tgccctgcgt aagcgggtgt gggcggacaa taaagtctta aactgaacaa aatagatcta
     3661 aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt
     3721 atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga
     3781 agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa gggcatgtaa agactatatt
     3841 cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa
     3901 cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc
     3961 ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca
     4021 cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc
     4081 cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct
     4141 gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg
     4201 cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct
     4261 gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc
     4321 gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc
     4381 cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca
     4441 tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat
     4501 gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg
     4561 ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac
     4621 ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt
     4681 ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgccat a
//
