LOCUS       sequence-275989-        5287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, 
            PI3KC2gamma-PX (1200-1310).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5287)
  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 5287)
  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: PI3KC2gamma-PX (1200-1310)
FEATURES             Location/Qualifiers
     source          1..5287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..676
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     634..656
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             683..700
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1102..1124)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1269..1287)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1355..1459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1460..1528,1529..2320)
                     /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(1678..1697)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2491..3079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2980..2999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3233..3250
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3323..3400
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3401..4483
                     /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(3420..3439)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4532..4537,4538..4555,4556..4562)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4570..4586
                     /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     4594..4610
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4594..4610
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4606..5133
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4626..4643)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4626..4642)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4635..4657)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5235..5240,5241..5256,5257..5263)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5271..5287
                     /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)."
ORIGIN
        1 tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct
       61 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
      121 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
      181 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
      241 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
      301 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
      361 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
      421 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
      481 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
      541 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
      601 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
      661 cgaccatcct ccaaaatcgg atctggttcc gcgtggatcc tcaaccaccc gcagcatcga
      721 acgtgccacc attctgggct tctcaaaaaa gtcctcaaac ctgtatctga ttcaagttac
      781 ccactctaac aatgaaacct cactgacgga aaaatcgttt gaacaattca gcaagctgca
      841 ttctcagctg caaaaacagt ttgcgtccct gaccctgccg gaatttccgc attggtggca
      901 cctgccgttt acgaacagcg atcatcgtcg cttccgtgac ctgaatcact atatggaaca
      961 aattctgaac gtgagccacg aagttaccaa ttctgattgc gtcctgagct ttttcctgtc
     1021 cgaagccgtt cagtaactcg agcggccgca tcgtgactga ctgacgatct gcctcgcgcg
     1081 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     1141 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1201 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
     1261 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     1321 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1381 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1441 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1501 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1561 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1621 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1681 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     1741 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1801 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1861 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1921 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1981 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2041 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2101 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2161 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2221 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2281 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2341 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2401 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2461 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2521 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2581 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2641 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2701 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2761 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2821 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2881 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2941 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3001 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3061 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3121 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3181 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3241 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3301 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
     3361 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     3421 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     3481 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     3541 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     3601 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     3661 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     3721 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     3781 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     3841 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     3901 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     3961 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     4021 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     4081 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     4141 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     4201 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     4261 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     4321 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     4381 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4441 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4501 taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4561 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4621 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4681 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4741 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
     4801 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
     4861 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
     4921 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
     4981 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
     5041 ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     5101 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     5161 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     5221 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
     5281 ataacaa
//
