LOCUS       sequence-276171-        5314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, mSNX13-PX 
            (558-677).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5314)
  AUTHORS   Chandra M, Chin YK, Mas C, Feathers JR, Paul B, Datta S, Chen KE, 
            Jia X, Yang Z, Norwood SJ, Mohanty B, Bugarcic A, Teasdale RD, Henne
            WM, Mobli M, Collins BM
  TITLE     Classification of the human phox homology (PX) domains based on 
            their phosphoinositide binding specificities.
  JOURNAL   Nat Commun. 2019 Apr 4;10(1):1528. doi: 10.1038/s41467-019-09355-y.
  PUBMED    30948714
REFERENCE   2  (bases 1 to 5314)
  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: "Nat 
            Commun."; date: "2019-04-4"; volume: "10(1):1528. doi"; pages: " 
            10.1038/s41467-019-09355-y"
COMMENT     Sequence Label: mSNX13-PX (558-677)
FEATURES             Location/Qualifiers
     source          1..5314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     44..66
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(495..512)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(519..1172)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(539..561)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    1195..1211
                     /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(1219..1225,1226..1241,1242..1247))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(1349..1876)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     1825..1847
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1839..1856
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1840..1856
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1872..1888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1872..1888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1896..1912
                     /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(1920..1926,1927..1944,1945..1950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             complement(1999..3081)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3043..3062
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3082..3159)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(3232..3249)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3403..3991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3483..3502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4162..4953,4954..5022))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4785..4804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5023..5127)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5195..5213
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
       61 cagaggtttt caccgtcatc accgaaacgc gcgaggcaga tcgtcagtca gtcacgatgc
      121 ggccgctcga gttaggcttt attttccaga aagtcataaa cacaatgtgc cagtgccgga
      181 cttgctttca tcatttccgg tgtcagcagc agctgcagat atgcattcag atcttttttg
      241 cgtttttcca gaaaatcgcg atccatatta ttaaaggttt ttttaccagg cagtttcaga
      301 atgctgctca gattttcaaa ctgttcggta atgcgcatgt gaaaatcatg aaaatcgcta
      361 taacggcgat aggttttcca catttcttcg gtattcagat tacgacgatg aacggtaatg
      421 gcatacagtg cataggtttt accatgatca ttgcacacac cggtatctga aatatatgca
      481 tgcagctgaa cgctggatcc acgcggaacc agatccgatt ttggaggatg gtcgccacca
      541 ccaaacgtgg cttgccagcc ctgcaaaggc catgctatat acttgctgga tttcaagtac
      601 ttatcaattt gtgggatagc ttcaatacgt tttttaaaac aaactaattt tgggaacgca
      661 tccaggcaca ttgggtccat gtataaaaca acatcaagag cgtcatacaa catgaagtca
      721 ggatgggtta catgatcacc atttaaatat gttttatgac ataaacgatc ttcgaacatt
      781 ttcagcattt caggtagctt gctaagaaaa tcaactttga gagtttcaaa gtctttacta
      841 tatgcaattc tcgaaacacc gtatctaata tccaaaaccg ctccttcaag cattgaaatc
      901 tctgcacgct cttttggaca accacccaac atgttgtgct tgtcagctat ataacgtatg
      961 atggccatag actgtgttaa tttaacatca ccatcaatat aataaggaag attgggaaac
     1021 tccaaaccca attcaaactt tttgtttcgc catttatcac cttcatcgcg ctcatacaaa
     1081 tgctcttcat atttttcttc aagatattcc aaaagaagtc gagtgggttg cacaaggccc
     1141 ttaattttcc aataacctag tataggggac atgaatactg tttcctgtgt gaaattgtta
     1201 tccgctcaca attccacaca ttatacgagc cgatgattaa ttgtcaacag ctcatttcag
     1261 aatatttgcc agaaccgtta tgatgtcggc gcaaaaaaca ttatccagaa cgggagtgcg
     1321 ccttgagcga cacgaattat gcagtgattt acgacctgca cagccatacc acagcttccg
     1381 atggctgcct gacgccagaa gcattggtgc accgtgcagt cgataagcta attccaacgc
     1441 catcaaaaat aattcgcgtc tggccttcct gtagccagct ttcatcaaca ttaaatgtga
     1501 gcgagtaaca acccgtcgga ttctccgtgg gaacaaacgg cggattgacc gtaatgggat
     1561 aggtcacgtt ggtgtagatg ggcgcatcgt aaccgtgcat ctgccagttt gaggggacga
     1621 cgacagtatc ggcctcagga agatcgcact ccagccagct ttccggcacc gcttctggtg
     1681 ccggaaacca ggcaaagcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     1741 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     1801 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     1861 ccgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactt
     1981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2041 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     2101 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     2161 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     2221 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     2281 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     2341 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     2401 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     2461 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     2521 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     2581 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     2641 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     2701 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     2761 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     2821 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     2881 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     2941 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     3001 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     3061 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     3121 tgccataccg cgaaaggttt tgcgccattc gatggtgtcg gaatttatgc ggtgtgaaat
     3181 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     3241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3841 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4081 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4441 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     4501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     4801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5161 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5221 tcttcaagaa ttatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     5281 cccgccaaca cccgctgacg cgccctgacg ggct
//
