LOCUS       sequence-259516-        5804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human TBC1D7 (19-293) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5804)
  AUTHORS   Qin J, Wang Z, Hoogeveen-Westerveld M, Shen G, Gong W, Nellist M, Xu
            W
  TITLE     Structural Basis of the Interaction between Tuberous Sclerosis 
            Complex 1 (TSC1) and Tre2-Bub2-Cdc16 Domain Family Member 7 
            (TBC1D7).
  JOURNAL   J Biol Chem. 2016 Apr 15;291(16):8591-601. doi: 
            10.1074/jbc.M115.701870. Epub 2016 Feb 18.
  PUBMED    26893383
REFERENCE   2  (bases 1 to 5804)
  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"; doi: 
            "10.1074/jbc.M115.701870"; journalName: "J Biol Chem"; date: 
            "2016-04-15- 15"; volume: "291"; issue: "16"; pages: "8591-601"
COMMENT     Sequence Label: pGEX-4T1 human TBC1D7 (19-293)
FEATURES             Location/Qualifiers
     source          1..5804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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             47..700
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     658..680
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             707..724
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             734..754
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1643..1665)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1810..1828)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1896..2000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2001..2069,2070..2861)
                     /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(2219..2238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3032..3620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3521..3540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3774..3791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3864..3941
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3942..5024
                     /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(3961..3980)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5073..5078,5079..5096,5097..5103)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5111..5127
                     /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     5135..5151
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5135..5151
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5147..5674
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5167..5184)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5167..5183)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5176..5198)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5776..5781,5782..5797,5798..5804)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact
       61 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
      121 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
      181 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
      241 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
      301 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
      361 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
      421 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
      481 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
      541 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
      601 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
      661 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg
      721 atccccggaa ttcgaaaacc tttacttcca gggctttcgt ggagttgaag aaaagaaatc
      781 attagaaatt ctcctaaaag atgaccgtct ggatactgag aaactttgta cttttagtca
      841 gaggttccct ctcccgtcca tgtaccgtgc attggtatgg aaggtgcttc taggaatctt
      901 gcctccacac cacgagtccc atgccaaggt gatgatgtat cgtaaggagc agtacttgga
      961 tgtccttcat gccctgaaag tcgttcgctt tgttagtgat gccacacctc aggctgaagt
     1021 ctatctccgc atgtatcagc tggagtctgg gaagttacct cgaagtccct cttttccact
     1081 ggagccagat gatgaagtgt ttcttgccat agctaaagcc atggaggaaa tggtggaaga
     1141 tagtgtcgac tgttactgga tcacccgacg ctttgtgaac caattaaata ccaagtaccg
     1201 ggattccttg ccccagttgc caaaagcgtt tgaacaatac ttgaatctgg aagatggcag
     1261 actgctgact catctgagga tgtgttccgc ggcgcccaaa cttccttatg atctctggtt
     1321 caagaggtgc tttgcgggat gtttgcctga atccagttta cagagggttt gggataaagt
     1381 tgtgagtgga tcctgtaaga tcctagtttt tgtagctgtc gaaattttat taacctttaa
     1441 aataaaagtt atggcactga acagtgcaga gaagataaca aagtttctgg aaaatattcc
     1501 ccaggacagc tcagacgcga tcgtgagcaa ggccattgac ttgtggcaca aacactgtgg
     1561 gaccccggtc cattcaagct gagcggccgc atcgtgactg actgacgatc tgcctcgcgc
     1621 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1681 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     1741 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa
     1801 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1861 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2221 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     2281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2521 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3721 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3781 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3841 gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg
     3901 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     3961 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     4021 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     4081 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     4141 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     4201 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     4261 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     4321 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     4381 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     4441 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     4501 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     4561 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     4621 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     4681 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     4741 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     4801 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     4861 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     4921 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4981 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5041 gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5101 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5161 ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     5221 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     5281 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
     5341 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
     5401 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
     5461 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
     5521 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
     5581 tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
     5641 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
     5701 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
     5761 ttctgaaatg agctgttgac aattaatcat cggctcgtat aatg
//
