LOCUS       sequence-276101-        5290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, IRAS-PX 
            (13-124).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5290)
  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 5290)
  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: IRAS-PX (13-124)
FEATURES             Location/Qualifiers
     source          1..5290
                     /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(1105..1127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1272..1290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1358..1462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1463..1531,1532..2323)
                     /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(1681..1700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2494..3082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2983..3002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3236..3253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3326..3403
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3404..4486
                     /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(3423..3442)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4535..4540,4541..4558,4559..4565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4573..4589
                     /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     4597..4613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4597..4613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4609..5136
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4629..4646)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4629..4645)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4638..4660)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5238..5243,5244..5259,5260..5266)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5274..5290
                     /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 gcagaaccgg ctaaggaagc
      721 tcgcgttgtg ggctcagaac tggtcgatac ctacaccgtc tacattattc aagtcaccga
      781 tggcagccat gaatggaccg ttaaacaccg ttatagtgat tttcatgacc tgcacgaaaa
      841 gctggtcgca gaacgcaaaa ttgataagaa cctgctgccg ccgaaaaaga ttatcggcaa
      901 aaatagccgt tctctggtcg aaaaacgcga aaaggacctg gaagtgtatc tgcagaaact
      961 gctggcagca tttccgggtg tgacgccgcg tgttctggca catttcctgc attttcactt
     1021 ctacgaaatc aacggttaac tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc
     1081 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     1141 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     1201 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt ataattcttg
     1261 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     1321 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1381 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1441 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1501 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1561 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1621 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1681 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     1741 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1801 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1861 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1921 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1981 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     2041 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2101 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2161 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2221 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2281 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2641 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3181 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3241 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     3301 cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat
     3361 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     3421 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     3481 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     3541 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     3601 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     3661 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     3721 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     3781 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     3841 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     3901 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     3961 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     4021 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     4081 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     4141 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     4201 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     4261 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     4321 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     4381 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     4441 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     4501 atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     4561 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     4621 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4681 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4741 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
     4801 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
     4861 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg
     4921 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     4981 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     5041 gatggcgttg gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     5101 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     5161 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     5221 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     5281 cggataacaa
//
