LOCUS       sequence-380473-        3184 bp    DNA     circular SYN 11-DEC-2025
DEFINITION  Expresses nuclease A-H124L from Staphylococcus aureus, with an A112C
            mutation, fused through a flexible linker to the transmembrane 
            domain of poly-Leu-1A32 and a His-tag, for fluorescent studies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3184)
  AUTHORS   Diaz Vazquez G, Cui Q, Senes A
  TITLE     Thermodynamic analysis of the GAS(right) transmembrane motif 
            supports energetic model of dimerization.
  JOURNAL   Biophys J. 2023 Jan 3;122(1):143-155. doi: 
            10.1016/j.bpj.2022.11.018. Epub 2022 Nov 12.
  PUBMED    36371634
REFERENCE   2  (bases 1 to 3184)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bpj.2022.11.018"; journalName: "Biophys J"; date: 
            "2023-01-3- 3"; volume: "122"; issue: "1"; pages: "143-155"
COMMENT     Sequence Label: pSN-A112C-1A32
FEATURES             Location/Qualifiers
     source          1..3184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     129..151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     143..160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     144..160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(231..355)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(900..995)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1032..1051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1033..1051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1056..1072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1056..1072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1174..1193)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1185..1994
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1252..1271)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2141..2729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2630..2649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2883..2900
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3059..3086)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(3178..3184,1..80))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
       61 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
      121 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag
      181 ctctagagct cgaattctac aggtcactaa taccatctaa gtagttggtt catagtgact
      241 gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
      301 tattgatatt tatatcattt tacgtttctc gttcaacttt cttgtacaaa gtggtgacta
      361 gcggatacaa ggacgggaca cctgaagagg aatacgctca ctcgccgttt atcaggaaac
      421 aggggaacat ttacaaacca aacaataaag aaatggataa cgacagcatc aacgaaaaga
      481 cacttcagga tgtccacacc aaggagattg acctggtcaa ccgagaccca aagcatttaa
      541 atgacgatgt ggtgaaggtg gactttgagg acgtgatcgc cgagcctgcc ggcacctaca
      601 gcttcgacgg cgtgtggaag gcgagcttca ccaccttcac agtaaccaaa tattggtgct
      661 acaggctgct gacagcgctg gtgggcatcc cactcgccct ggtatggggc atcttcttcg
      721 ccatcctctc cttcatccac atctgggccg tggtgccttg cgtgaagagc tacctaatcg
      781 agatccactg catcagtcga gtttactcca tctgtgtgca caccttctgc gacccactct
      841 ttgaggccat ggggaaatgc tttagtaacg tccgggtcac tgctactaag gtggtgtaac
      901 aactttatta tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga
      961 acaggtcact atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg
     1021 gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1081 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     1141 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     1201 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1261 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1321 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1381 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1441 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1501 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1561 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1621 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1681 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     1741 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1801 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1861 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1921 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1981 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     2041 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     2101 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2161 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2221 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2281 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2341 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2401 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2461 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2521 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2581 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2641 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2701 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2761 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2821 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2881 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2941 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3001 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     3061 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     3121 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     3181 tgtc
//
