LOCUS       sequence-127069-        5972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse Tropomodulin3, 1-325 domain for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5972)
  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 5972)
  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-mTmod3-1-325_domain
FEATURES             Location/Qualifiers
     source          1..5972
                     /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(2026..2048)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2193..2211)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2279..2383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2384..2452,2453..3244)
                     /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(2602..2621)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3415..4003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3904..3923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4157..4174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4247..4324
                     /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             4325..5407
                     /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(4344..4363)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5420..5441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5456..5461,5462..5479,5480..5486)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5494..5510
                     /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     5518..5534
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5518..5534
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5530..5972,1..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(5550..5567)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5550..5566)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5559..5581)
                     /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 gcagcagtag cgtttcgagc tcaagcttaa ttcatcgtga
     1981 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2041 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2101 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     2161 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     2221 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2281 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2341 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2401 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2461 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2521 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2581 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     2641 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2701 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2761 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2821 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2881 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     2941 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3001 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3061 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3121 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3181 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3241 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3301 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3361 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3541 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4081 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4141 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4201 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     4261 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     4321 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     4381 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     4441 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     4501 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     4561 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     4621 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     4681 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     4741 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     4801 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     4861 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     4921 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     4981 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     5041 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     5101 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     5161 ctcatgttat atcccgccgt caaccaccat caaacaggat tttcgcctgc tggggcaaac
     5221 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     5281 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     5341 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5401 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5461 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5521 gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg
     5581 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5641 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5701 gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc
     5761 gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat
     5821 gcgcccatct acaccaacgt aacctatccc attacggtca atccgccgtt tgttcccacg
     5881 gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa
     5941 ggccagacgc gaattatttt tgatggcgtt gg
//
