LOCUS       sequence-126669-        6053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse Tropomodulin3, WT for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6053)
  AUTHORS   Lim CY, Bi X, Wu D, Kim JB, Gunning PW, Hong W, Han W
  TITLE     Tropomodulin3 is a novel Akt2 effector regulating insulin-stimulated
            GLUT4 exocytosis through cortical actin remodeling.
  JOURNAL   Nat Commun. 2015 Jan 9;6:5951. doi: 10.1038/ncomms6951.
  PUBMED    25575350
REFERENCE   2  (bases 1 to 6053)
  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: "2015-01-9"; volume: "6:5951. doi"; pages: " 
            10.1038/ncomms6951"
COMMENT     Sequence Label: pGEX-KG-mTmod3-WT
FEATURES             Location/Qualifiers
     source          1..6053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(187..192,193..208,209..215)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    223..239
                     /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             262..915
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     873..895
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             922..939
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2107..2129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2274..2292)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2360..2464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2465..2533,2534..3325)
                     /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(2683..2702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3985..4004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4238..4255
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4328..4405
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4406..5488
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4425..4444)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5501..5522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5537..5542,5543..5560,5561..5567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5575..5591
                     /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     5599..5615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5599..5615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5611..6053,1..85)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5631..5648)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5631..5647)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5640..5662)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aattacgtta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
       61 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
      121 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
      181 gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat
      241 ttcacacagg aaacagtatt catgtcccct atactaggtt attggaaaat taagggcctt
      301 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
      361 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
      421 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
      481 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
      541 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
      601 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
      661 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
      721 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
      781 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
      841 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
      901 gaccatcctc caaaatcgga tctggttccg cgtggatccc cgggaatttc cggtggtggt
      961 ggtggaattc agatggcact gccgttccgg aaggacttgg gggactacaa ggacctggat
     1021 gaggacgagc tccttggcaa gctgtccgaa tcagagctga aacagctgga gactgtcctg
     1081 gatgatcttg accccgagaa tgcccttctg cctgcagggt ttcggcagaa gaaccagaca
     1141 tcaaagtctg ccacgggacc ctttgataga gaacggctcc tttcttacct agagaagcaa
     1201 gcattggagc acaaagaccg ggacgattat gtgccctaca ctggagagaa gaaagggaaa
     1261 atttttatcc ccaaacagaa gccagcgcag actcttacag aagaaactat atccctcgac
     1321 ccagagttag aagaagcttt gacaagtgct tcagacacag agctgtgcga ccttgcagct
     1381 attcttggga tgcacaatct gattgcagac acaccgttct gtgatgtgct gggaagtagt
     1441 aacggtgtca accaagagcg ttttcccaat gtggttaaag gggaaaagat tcttccagtg
     1501 tttgatgagc caccaaatcc aaccaacgtt gaagagagtt taaagagaat tagggaaaac
     1561 gatgcccgtc ttgttgaagt taatttgaat aatataaaaa atatcccaat tccaactcta
     1621 aaagattttg caaagacttt ggaagccaac acgcatgtga aacatttcag tctcgcggcc
     1681 accaggagca atgaccccgt ggctgttgct tttgcagata tgctgaaagt gaacaaaact
     1741 ttgaagagct taaatatgga gtccaacttc atcacaggcg caggggtttt agcactgatt
     1801 gatgctttga gagataatga gaccctgatg gagctcaaga ttgacaatca gagacagcag
     1861 ttgggcacct cagtagaatt agaaatggcc aagatgcttg aggaaaatac aaatatcctt
     1921 aaatttggat atcagttcac gcagcaggga ccccgaacca gggcagccaa cgcgataacg
     1981 aagaacaatg acttagtgcg caagaggcga attgaaggag accaccagta acgtttcgag
     2041 ctcaagctta attcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg
     2101 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2161 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     2221 catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt
     2281 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2341 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2401 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2461 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2521 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2581 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2641 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2701 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2761 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2821 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2881 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2941 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3001 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3061 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3121 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3181 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3241 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3301 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4201 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4261 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4321 aaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
     4381 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     4441 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     4501 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     4561 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     4621 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     4681 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     4741 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     4801 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     4861 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     4921 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     4981 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     5041 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     5101 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     5161 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     5221 atacgacgat accgaagaca gctcatgtta tatcccgccg tcaaccacca tcaaacagga
     5281 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     5341 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     5401 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5461 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     5521 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     5581 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
     5641 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5701 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5761 cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg
     5821 ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac
     5881 tggcagatgc acggttacga tgcgcccatc tacaccaacg taacctatcc cattacggtc
     5941 aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt
     6001 gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt tgg
//
