LOCUS       sequence-275986-        5374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, 
            PI3KC2alpha-PX (1405-1545).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5374)
  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 5374)
  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: PI3KC2alpha-PX (1405-1545)
FEATURES             Location/Qualifiers
     source          1..5374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1242..1264)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1409..1427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1495..1599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1600..1668,1669..2460)
                     /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(1818..1837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2631..3219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3120..3139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3373..3390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3463..3540
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3541..4623
                     /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(3560..3579)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4672..4677,4678..4695,4696..4702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4710..4726
                     /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     4734..4750
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4734..4750
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4746..5273
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4766..4783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4766..4782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4775..4797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccgacgaac cgattctgtc attttctccg
      781 aagacctact cattccgtca agatggccgc attaaagaag tgtccgtttt cacctatcat
      841 aagaagtaca acccggataa gcattacatc tacgtggttc gtatcctgcg cgaaggccag
      901 attgaaccga gttttgtgtt ccgtaccttt gacgaatttc aagaactgca taacaaactg
      961 tccattatct ttccgctgtg gaagctgccg ggcttcccga atcgtatggt tctgggtcgc
     1021 acgcacatta aagatgtcgc ggccaagcgt aagatcgaac tgaacagcta cctgcagtct
     1081 ctgatgaatg cgagcaccga tgtcgcagaa tgcgacctgg tgtgtacgtt tttccacccg
     1141 ctgctgcgcg acgaaaaggc agaaggtatc gcttaactcg agcggccgca tcgtgactga
     1201 ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1261 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1321 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     1381 ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1441 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1561 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1621 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1681 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1741 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1801 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     1861 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1921 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1981 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2041 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2101 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     2161 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2221 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2281 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2341 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2761 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3241 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3301 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3361 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3421 ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc
     3481 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     3541 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     3601 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     3661 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     3721 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     3781 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     3841 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     3901 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     3961 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     4021 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     4081 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     4141 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     4201 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     4261 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     4321 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     4381 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     4441 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     4501 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     4561 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4621 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
     4681 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     4741 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     4801 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4861 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4921 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
     4981 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
     5041 ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
     5101 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
     5161 agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg
     5221 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
     5281 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
     5341 ataacggttc tggcaaatat tctgaaatga gctg
//
