LOCUS       sequence-191842-        6918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            ARHGEF26-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  AUTHORS   Teyra J, Huang H, Jain S, Guan X, Dong A, Liu Y, Tempel W, Min J, 
            Tong Y, Kim PM, Bader GD, Sidhu SS
  TITLE     Comprehensive Analysis of the Human SH3 Domain Family Reveals a Wide
            Variety of Non-canonical Specificities.
  JOURNAL   Structure. 2017 Oct 3;25(10):1598-1610.e3. doi: 
            10.1016/j.str.2017.07.017. Epub 2017 Sep 7.
  PUBMED    28890361
REFERENCE   2  (bases 1 to 6918)
  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: 
            "10.1016/j.str.2017.07.017"; journalName: "Structure"; date: 
            "2017-10-3- 3"; volume: "25"; issue: "10"; pages: "1598-1610.e3"
COMMENT     Sequence Label: pHH0103_ARHGEF26-1/1
FEATURES             Location/Qualifiers
     source          1..6918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..107
                     /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             108..1190
                     /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(127..146)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1433..1438,1439..1454,1455..1461)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1469..1485
                     /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     1474..1496
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1570..1587
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1603..2256
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2214..2236
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2263..2286
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2299..2316
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2545..2592
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4185..4640)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4322..4343)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4534..4553
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4682..4704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4790..4930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4845..4864
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4945..4962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5116..5703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5874..6665,6666..6734))
                     /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     6497..6516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6735..6839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(6907..6918,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtcttca agaattcccg acaccatcga atggtgcaaa acctttcgcg
       61 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
      121 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
      181 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
      241 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
      301 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
      361 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
      421 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
      481 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
      541 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
      601 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
      661 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
      721 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
      781 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
      841 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
      901 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
      961 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     1021 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     1081 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     1141 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     1201 attaatgtaa gttagctcac tcattaggca caattctcat gtttgacagc ttatcatcga
     1261 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1321 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1381 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1441 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1501 agccagtccg tttaggtgtt ttcacgagca cttcaccaac aaggaccata gattatgaaa
     1561 atcgaagaac accatcacca tcaccattcc agcggtaagc ttatgtcccc tatactaggt
     1621 tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa
     1681 aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt
     1741 gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca
     1801 cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca
     1861 aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt
     1921 tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta
     1981 cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat
     2041 catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac
     2101 ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc
     2161 ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg
     2221 caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag
     2281 gggcccctgt ccagcggtct ggttccgcgt ggttccggta ccgcggccca gccggccatg
     2341 gcccagggcg cgctgaccca ggttgaaatt gtccgttcct tcaccgcaaa acagccagat
     2401 gaactgtctc tgcaagttgc cgatgtcgtt ctgatttacc agcgtgtttc cgatggttgg
     2461 tacgaaggtg aacgtctgcg tgatggtgaa cgtggttggt ttccaatgga atgtgccaaa
     2521 gaaatcaccg cggccgcata ataaaccgat acaattaaag gctccttttg gagccttttt
     2581 tttggagatt ttcaacgtga aaaaattatt attcgcaatt cctttagttg ttcctttcta
     2641 ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa ccccatacag aaaattcatt
     2701 tactaacgtc tggaaagacg acaaaacttt agatcgttac gctaactatg agggctgtct
     2761 gtggaatgct acaggcgttg tagtttgtac tggtgacgaa actcagtgtc tagctagagt
     2821 ggcggtggct ctggttccgg tgattttgat tatgaaaaga tggcaaacgc taataagggg
     2881 gctatgaccg aaaatgccga tgaaaacgcg ctacagtctg acgctaaagg caaacttgat
     2941 tctgtcgcta ctgattacgg tgctgctatc gatggtttca ttggtgacgt ttccggcctt
     3001 gctaatggta atggtgctac tggtgatttt gctggctcta attcccaaat ggctcaagtc
     3061 ggtgacggtg ataattcacc tttaatgaat aatttccgtc aatatttacc ttccctccct
     3121 caatcggttg aatgtcgccc ttttgtcttt ggcgctggta aaccatatga attttctatt
     3181 gattgtgaca aaataaactt attccgtggt gtctttgcgt ttcttttata tgttgccacc
     3241 tttatgtatg tattttctac gtttgctaac atactgcgta ataaggagtc ttaatcatgc
     3301 cagttctttt ggctagcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
     3361 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
     3421 cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc
     3481 ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg
     3541 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     3601 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     3661 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     3721 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     3781 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     3841 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     3901 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     3961 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     4021 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     4081 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     4141 atcgcttcac gaccacgctg atgagcttta ccgcaggatc cgcgaaattg taagcgttaa
     4201 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4261 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4321 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4381 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4441 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4501 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4561 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4621 tgcgccgcta cagggcgcgt cgcggatcct gcctcgcgcg tttcggtgat gacggtgaaa
     4681 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4741 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     4801 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     4861 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4921 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4981 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5041 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5101 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5161 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccagggtt tccccctgga
     5221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5521 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6181 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6241 ggttcccaac gatcaaggcg agttacatga tcccccatat tgtgcaaaaa agcggttagc
     6301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6481 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6841 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6901 tataaaaata ggcgtatc
//
