LOCUS       sequence-276013-        5329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, PXK-PX 
            (1-125).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  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 5329)
  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: PXK-PX (1-125)
FEATURES             Location/Qualifiers
     source          1..5329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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)."
     CDS             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1161..1183)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1328..1346)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1414..1518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1519..1587,1588..2379)
                     /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     complement(1737..1756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2550..3138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3039..3058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3292..3309
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3382..3459
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3460..4542
                     /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     complement(3479..3498)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4591..4596,4597..4614,4615..4621)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4645
                     /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)."
     primer_bind     4653..4669
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4653..4669
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4665..5192
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4685..4702)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4685..4701)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4694..4716)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5294..5299,5300..5315,5316..5322)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccatg
      721 gcgttcatgg agaagccgcc ggcgggtaaa gtgctgctgg acgataccgt tccgctgacc
      781 gcggcgattg aggcgagcca gagcctgcaa agccacaccg aatacatcat tcgtgtgcag
      841 cgtggcatca gcgttgaaaa cagctggcaa attgtgcgtc gttatagcga cttcgatctg
      901 ctgaacaaca gcctgcagat tgcgggtctg agcctgccgc tgccgccgaa gaaactgatc
      961 ggcaacatgg accgtgagtt tattgcggaa cgtcagaagg gtctgcaaaa ctacctgaac
     1021 gtgatcacca ccaaccacat tctgagcaac tgcgagctgg ttaagaaatt tctggacccg
     1081 aacaactata gctaactcga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt
     1141 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     1201 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1261 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga
     1321 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1381 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1441 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1501 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1621 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1681 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1741 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1801 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1921 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2041 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     2101 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2341 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2401 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3241 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3301 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3361 atttcacacc gcataaattc cgacaccatc gaatggcgca aaacctttcg cggtatggca
     3421 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     3481 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     3541 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     3601 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     3661 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     3721 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     3781 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     3841 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     3901 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     3961 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     4021 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     4081 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     4141 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     4201 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     4261 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     4321 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     4381 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     4441 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4501 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4561 aagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4621 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg
     4681 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     4741 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     4801 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
     4861 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg
     4921 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct
     4981 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc
     5041 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg
     5101 gcgttggaat tagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat
     5161 cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg
     5221 cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt
     5281 ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaa
//
