LOCUS       sequence-498089-        5692 bp    DNA     circular SYN 23-JAN-2026
DEFINITION  Bacterial expression of gamma 14-3-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5692)
  AUTHORS   Espejo AB, Gao G, Black K, Gayatri S, Veland N, Kim J, Chen T, Sudol
            M, Walker C, Bedford MT
  TITLE     PRMT5 C-terminal Phosphorylation Modulates a 14-3-3/PDZ Interaction 
            Switch.
  JOURNAL   J Biol Chem. 2017 Feb 10;292(6):2255-2265. doi: 
            10.1074/jbc.M116.760330. Epub 2016 Dec 28.
  PUBMED    28031468
REFERENCE   2  (bases 1 to 5692)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 23, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M116.760330"; journalName: "J Biol Chem"; date: 
            "2017-02-10- 10"; volume: "292"; issue: "6"; pages: "2255-2265"
COMMENT     Sequence Label: pGEX-gamma-14-3-3
FEATURES             Location/Qualifiers
     source          1..5692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        187..264
                     /gene="lacI"
                     /label=lacI promoter
     CDS             265..1347
                     /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(284..303)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1396..1401,1402..1419,1420..1426)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1434..1450
                     /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     1458..1474
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1458..1474
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1470..1997
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1490..1507)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1490..1506)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1499..1521)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2099..2104,2105..2120,2121..2127)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2135..2151
                     /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             2174..2827
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2785..2807
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2834..2851
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3658..3680)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3825..3843)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3911..4015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4016..4084,4085..4876)
                     /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(4234..4253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5047..5635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5536..5555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
       61 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      121 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      181 aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
      241 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      301 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      361 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      421 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      481 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      541 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      601 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      661 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      721 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      781 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      841 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      901 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      961 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     1021 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     1081 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     1141 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1201 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1261 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1321 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     1381 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1441 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
     1501 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     1561 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1621 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
     1681 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
     1741 ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca
     1801 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
     1861 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaattagct
     1921 tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat
     1981 ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg
     2041 gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt
     2101 gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     2161 ggaaacagta ttcatgtccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc
     2221 cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga
     2281 tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc
     2341 ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc
     2401 tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga
     2461 aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga
     2521 aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg
     2581 tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta
     2641 tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt
     2701 agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact tgaaatccag
     2761 caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg gcgaccatcc
     2821 tccaaaatcg gatctggttc cgcgtggatc catggttgac cgtgaacagc tggttcagaa
     2881 agcgcgcctg gcggagcagg ctgagcgcta cgacgacatg gctgcggcga tgaaaaacgt
     2941 gactgaactg aacgaaccgc tgagcaacga agagcgtaac ctgctgtctg ttgcgtacaa
     3001 gaacgttgtg ggtgcgcgtc gttctagctg gcgtgttatc tcttctatcg aacagaaaac
     3061 cagcgcggac ggtaacgaaa agaaaatcga aatggttcgt gcgtatcgtg aaaagatcga
     3121 aaaagagctg gaagcggttt gccaggatgt tctgtccctg ctggacaact atctgatcaa
     3181 aaactgttct gaaacccagt acgaatctaa agttttttac ctgaaaatga aaggtgacta
     3241 ctaccgttac ctggcagaag tggcgaccgg tgagaaacgc gcgaccgttg ttgaaagctc
     3301 tgaaaaagcg tactctgagg cgcatgaaat ctctaaagaa cacatgcagc cgacccaccc
     3361 gatccgtctg ggtctggcgc tgaactactc tgttttctat tacgaaatcc agaacgctcc
     3421 ggaacaggcg tgtcacctgg ctaaaaccgc gttcgacgac gctatcgcag aactggacac
     3481 cctgaacgag gattcctaca aagactctac cctgattatg cagctgctgc gtgacaacct
     3541 gaccctgtgg acttccgatc agcaggacga tgacggtggt gaaggtaaca acctcgagcg
     3601 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     3661 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3721 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     3781 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
     3841 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3901 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3961 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4021 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4081 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4141 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4201 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     4261 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4321 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4381 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4441 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4501 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     4561 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4621 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4681 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4741 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4801 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4861 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4921 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4981 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5041 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5101 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5161 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5221 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5281 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5341 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5401 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5461 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5521 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5581 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5641 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc ac
//
