LOCUS       sequence-275993-        5341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human phox homology (PX) domain, RICS-PX 
            (127-255).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5341)
  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 5341)
  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: RICS-PX (127-255)
FEATURES             Location/Qualifiers
     source          1..5341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             13..540
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(33..50)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(33..49)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(42..64)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(642..647,648..663,664..670)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    678..694
                     /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             717..1370
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1328..1350
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1377..1394
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1850..1872)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2017..2035)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2103..2207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2208..2276,2277..3068)
                     /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(2426..2445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3239..3827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3728..3747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3981..3998
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4071..4148
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4149..5231
                     /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(4168..4187)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5280..5285,5286..5303,5304..5310)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5318..5334
                     /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 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
       61 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
      121 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
      181 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
      241 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
      301 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
      361 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
      421 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
      481 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
      541 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
      601 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
      661 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt
      721 cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg
      781 aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc
      841 gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg
      901 atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt
      961 tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata
     1021 ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt
     1081 ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat
     1141 atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg
     1201 ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac
     1261 gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc
     1321 ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg
     1381 ttccgcgtgg atccaatgtc gaatttggta gcatccaact gagcctgagc gaagaacaga
     1441 acgaagtgat gaagaatggc tgcgaaagca aggaactggt ttatctggtt cagattgcct
     1501 gccaaggcaa atcctggatc gtgaagcgta gctatgaaga ttttcgcgtt ctggacaaac
     1561 atctgcacct gtgtatttac gatcgtcgct tcagtcagct gtccgaactg ccgcgtagtg
     1621 ataccctgaa ggactcaccg gaatcggtca cgcaaatgct gatggcatac ctgagccgcc
     1681 tgtctgcgat tgccggcaac aaaatcaatt gcggtccggc tctgacctgg atggaaatcg
     1741 acaacaaggg taatcatctg ctggtgcacg aagaaagctc ttaactcgag cggccgcatc
     1801 gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1861 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1921 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     1981 gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     2041 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     2101 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2161 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2221 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2281 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2341 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2401 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2461 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2521 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2581 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2641 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2701 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     2761 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2821 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2881 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2941 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3001 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3061 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3121 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3181 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3361 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3841 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3901 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3961 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4021 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg
     4081 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     4141 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     4201 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     4261 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     4321 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     4381 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     4441 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     4501 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     4561 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     4621 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     4681 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     4741 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     4801 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     4861 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     4921 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     4981 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     5041 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     5101 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     5161 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5221 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
     5281 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5341 a
//
