LOCUS       sequence-127066-        5486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse Tropomodulin3, 1-163 domain for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5486)
  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 5486)
  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_163_domain
FEATURES             Location/Qualifiers
     source          1..5486
                     /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(1540..1562)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1707..1725)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1793..1897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1898..1966,1967..2758)
                     /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(2116..2135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2929..3517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3418..3437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3671..3688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3761..3838
                     /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             3839..4921
                     /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(3858..3877)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    4934..4955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4970..4975,4976..4993,4994..5000)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5008..5024
                     /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     5032..5048
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5032..5048
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5044..5486,1..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(5064..5081)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5064..5080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5073..5095)
                     /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 ttagcgtttc gagctcaagc ttaattcatc gtgactgact
     1501 gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1561 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1621 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     1681 agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     1741 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1801 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1861 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1921 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1981 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2041 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2101 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     2161 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2221 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2281 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2341 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2401 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     2461 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2521 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2581 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2641 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2701 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2761 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2821 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2881 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2941 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3001 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3061 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3121 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3181 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3241 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3301 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3361 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3421 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3481 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3541 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3601 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3661 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3721 ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggtgcaa
     3781 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     3841 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     3901 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     3961 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     4021 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     4081 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     4141 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     4201 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     4261 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     4321 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     4381 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     4441 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     4501 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     4561 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     4621 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     4681 ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     4741 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     4801 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     4861 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     4921 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     4981 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5041 gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5101 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5161 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5221 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
     5281 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
     5341 atctacacca acgtaaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
     5401 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
     5461 acgcgaatta tttttgatgg cgttgg
//
