LOCUS       sequence-498085-        5689 bp    DNA     circular SYN 23-JAN-2026
DEFINITION  Bacterial expression of beta/alpha 14-3-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  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 5689)
  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-beta/alpha-14-3-3
FEATURES             Location/Qualifiers
     source          1..5689
                     /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(3655..3677)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3822..3840)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3908..4012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4013..4081,4082..4873)
                     /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(4231..4250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5044..5632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5533..5552
                     /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 catgaccatg gacaaatctg aactggttca
     2881 gaaagcgaaa ctggctgaac aggcggaacg ttacgacgac atggctgcgg caatgaaagc
     2941 ggttaccgaa cagggtcacg aactgtctaa cgaagagcgt aacctgctgt ctgttgcgta
     3001 caaaaacgtt gttggtgcgc gtcgttcttc ttggcgtgtt atctcttcta tcgaacagaa
     3061 gactgaacgt aacgaaaaaa agcagcagat gggtaaagag taccgtgaaa aaatcgaagc
     3121 agaactgcag gacatctgca acgacgttct ggaactgctg gacaagtatc tgatcccgaa
     3181 cgcgactcag ccggaatcta aagtgttcta cctgaaaatg aagggtgatt acttccgtta
     3241 cctgtctgaa gttgcgtctg gtgacaacaa acagaccacc gtttctaact ctcagcaggc
     3301 gtaccaggaa gcgtttgaaa tctctaaaaa agaaatgcag ccgacccacc cgatccgtct
     3361 gggtctggcg ctgaacttct ctgttttcta ttacgaaatc ctgaactctc cggaaaaagc
     3421 ttgctctctg gcgaaaactg cgttcgacga agctattgcg gaactggaca ctctgaacga
     3481 ggagtcttac aaagattcta ccctgatcat gcagctgctg cgtgataacc tgactctgtg
     3541 gacctctgaa aaccagggcg atgaaggtga cgcgggtgag ggtgaaaacc tcgagcggcc
     3601 gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     3661 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3721 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     3781 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     3841 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3901 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3961 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4021 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4081 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4141 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4201 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4261 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4321 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4381 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4441 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4501 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     4561 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4621 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4681 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4741 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4801 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4861 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4921 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4981 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5041 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5101 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5161 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5221 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5281 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5341 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5401 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5461 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5521 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5581 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5641 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcac
//
