LOCUS       sequence-256151-        7610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human INPP5A in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7610)
  AUTHORS   Erneux C, De Smedt F, Moreau C, Rider M, Communi D
  TITLE     Production of recombinant human brain type I 
            inositol-1,4,5-trisphosphate 5-phosphatase in Escherichia coli. Lack
            of phosphorylation by protein kinase C.
  JOURNAL   Eur J Biochem. 1995 Dec 1;234(2):598-602. doi: 
            10.1111/j.1432-1033.1995.598_b.x.
  PUBMED    8536709
REFERENCE   2  (bases 1 to 7610)
  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"; doi: 
            "10.1111/j.1432-1033.1995.598_b"; journalName: "Eur J Biochem"; 
            date: "1995-12-1- 1"; volume: "234"; issue: "2"; pages: "598-602"
COMMENT     Sequence Label: Human INPP5A
FEATURES             Location/Qualifiers
     source          1..7610
                     /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)."
     primer_bind     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             123..1223
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1080..1089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1197..1220
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1254..1265
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2793..2810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2793..2809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2802..2824)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2988..3005)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3158..3244
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3336..3363
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3382..3473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3474..3542,3543..4334)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3692..3711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4505..5093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4994..5013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5247..5264
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5279..5419
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5345..5364)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5505..5527
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5521..5712)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6208..6285
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6286..7368
                     /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(6305..6324)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaac tgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg
      181 gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc
      241 atccggataa actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca
      301 ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa
      361 tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt
      421 acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca
      481 aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac
      541 tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc
      601 cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta
      661 aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga
      721 ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata
      781 aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca
      841 aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg
      901 ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcgaaagagt
      961 tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc
     1021 tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg
     1081 ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt
     1141 tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg
     1201 aagccctgaa agacgcgcag actaattcga gctcggtacc gtcctctctc gtgatcgagg
     1261 gaaggatttc agaattcgga tccatggcgg ggaaggcggc cgccccgggc accgcggtgc
     1321 tgctggtcac ggccaacgtg ggctcgctct tcgacgaccc agaaaacctg cagaagaact
     1381 ggcttcggga attttaccag gtcgtgcaca cacacaagcc gcacttcatg gccttgcact
     1441 gtcaggagtt tggagggaag aactacgagg cctccatgtc ccacgtggac aagttcgtca
     1501 aagaactatt gtcgagtgat gcgatgaaag aatataacag ggctcgagtc tacctggatg
     1561 aaaactacaa atcccaggag cacttcacgg cactaggaag cttttatttt cttcatgagt
     1621 ccttaaaaaa catctaccag tttgacttta aagctaagaa gtatagaaag gtcgctggca
     1681 aagagatcta ctcggatacc ttagagagca cgcccatgct ggagaaggag aagtttccgc
     1741 aggactactt ccccgagtgc aaatggtcaa gaaaaggctt catccggacg aggtggtgca
     1801 ttgcagactg tgcctttgac ttggtgaata tccatctttt ccatgatgct tccaatctgg
     1861 tcgcctggga aacaagccct tccgtgtact cgggaatccg gcacaaggca ctgggctacg
     1921 tgctggacag aatcattgat cagcgattcg agaaggtttc ctactttgta tttggtgatt
     1981 tcaacttccg gctggattcc aagtccgtcg tggagacgct ctgcacaaaa gccaccatgc
     2041 agacggtccg ggccgccgac accaatgaag tggtgaagct catatttcgt gagtcggaca
     2101 acgaccggaa ggttatgctc cagttagaaa agaaactctt cgactacttc aaccaggagg
     2161 ttttccgaga caacaacggc accgcgctct tggagtttga caaggagttg tctgtcttta
     2221 aggacagact gtatgaactg gacatctcgt tccctcccag ctacccgtac agtgaggacg
     2281 cccgccaggg tgagcagtac atgaacaccc ggtgcccagc ctggtgtgac cgcatcctca
     2341 tgtccccgtc tgccaaggag ctggtgctgc ggtcggagag cgaggagaag gttgtcacct
     2401 atgaccacat tgggcccaac gtctgcatgg gagaccacaa gcccgtgttc ctggccttcc
     2461 gaatcatgcc cggggcaggt aaacctcatg cccatgtgca caagtgttgt gtcgtgcagt
     2521 gacgtggtgg gaagagatgc cagcgccacg agaggacact tcgtgagcct ccctgtagcc
     2581 gtagaccgaa tacgcactct tgaaagctgc atcgagaacc cgcccaagcg ccacctgcta
     2641 gacggccagc cccacacttc gcttcagcct ccggaccatt ccggagcagc ctcacatacc
     2701 tcactgtctc gtctgtctat gtgacattaa gtagaaatat tggaattcga tatcaagctt
     2761 atcgataccg tcgacctgca ggcaagcttg gcactggccg tcgttttaca acgtcgtgac
     2821 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2881 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2941 ggcgaatggc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     3001 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     3061 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     3121 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     3181 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     3241 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     3301 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     3361 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
     3421 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3481 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3541 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3601 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3661 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     3721 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3781 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3841 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3901 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3961 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4021 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4081 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4141 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4201 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4261 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4321 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4381 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4441 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4501 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4561 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4621 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4681 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4741 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4801 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4861 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4921 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4981 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5041 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5101 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5161 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5221 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5281 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     5341 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     5401 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5461 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5521 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     5581 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     5641 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     5701 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     5761 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     5821 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     5881 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     5941 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     6001 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     6061 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     6121 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     6181 ccaggaccca acgctgcccg aaattccgac accatcgaat ggcgcaaaac ctttcgcggt
     6241 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     6301 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     6361 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     6421 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     6481 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     6541 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     6601 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     6661 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     6721 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     6781 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     6841 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     6901 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     6961 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     7021 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     7081 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     7141 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     7201 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     7261 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7321 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     7381 taatgtaagt tagctcactc attaggcaca attctcatgt ttgacagctt atcatcgact
     7441 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     7501 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     7561 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
//
