LOCUS       sequence-275994-        5347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, mPLD2-PX 
            (62-192).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5347)
  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 5347)
  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: mPLD2-PX (62-192)
FEATURES             Location/Qualifiers
     source          1..5347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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             47..700
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     658..680
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             707..724
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1186..1208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1353..1371)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1439..1543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1544..1612,1613..2404)
                     /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(1762..1781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2575..3163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3064..3083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3317..3334
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3407..3484
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3485..4567
                     /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(3504..3523)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4616..4621,4622..4639,4640..4646)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4654..4670
                     /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     4678..4694
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4678..4694
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4690..5217
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4710..4727)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4710..4726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4719..4741)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5319..5324,5325..5340,5341..5347)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact
       61 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
      121 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
      181 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
      241 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
      301 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
      361 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
      421 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
      481 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
      541 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
      601 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
      661 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg
      721 atccggtgtt ccggttattg cacaggttgt tggcaccgaa cgttatacca gcggtagcaa
      781 agttggcacc tgtaccctgt atagcgttcg tctgacccat ggtgatttta cctggaccac
      841 caaaaaaaaa tttcgccatt ttcaggaact gcatcgcgat ctgcagcgtc ataaagttct
      901 gatgagcctg ctgccgctgg cacgttttgc agttacccat agtccggcac gtgaagcagc
      961 agcagaagat attccgagcc tgcctcgtgg tggtagcgaa ggtagcgcac gtcataccgc
     1021 aagcaaacag aaatatctgg aaaattatct gaatcgcctg ctgaccatga gcttttatcg
     1081 taattatcat gccatgaccg aatttctgga agtgagctaa ctcgagcggc cgcatcgtga
     1141 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     1201 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     1261 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     1321 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     1381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     1801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     2101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2701 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3241 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3301 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3361 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     3421 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     3481 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     3541 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     3601 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     3661 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     3721 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     3781 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     3841 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     3901 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     3961 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     4021 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     4081 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     4141 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     4201 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     4261 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     4321 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     4381 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     4441 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     4501 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4561 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccc caggctttac
     4621 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     4681 gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg
     4741 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4801 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4861 gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc
     4921 gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat
     4981 gcgcccatct acaccaacgt gacctatccc attacggtca atccgccgtt tgttcccacg
     5041 gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa
     5101 ggccagacgc gaattatttt tgatggcgtt ggaattagct tatcgactgc acggtgcacc
     5161 aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc
     5221 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga
     5281 catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg
     5341 tataatg
//
