LOCUS       sequence-213641-        7845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of GNA15_G-alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7845)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 7845)
  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: "Mol Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_GNA15_G-alpha
FEATURES             Location/Qualifiers
     source          1..7845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2040..2087
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3680..4135)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3817..3838)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4029..4048
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4177..4199)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4285..4425
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4340..4359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4440..4457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4611..5198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5369..6160,6161..6229))
                     /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     5992..6011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6230..6334)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6402..6420
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6443..6520
                     /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             6521..7603
                     /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(6540..6559)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggccgctcg ttctctgact tggcgttgtt gcccatggtg cctgaccgaa gacgagaaag
      961 ctgcggcccg tgttgaccaa gaaattaacc gtattctgct ggagcagaag aaacaggatc
     1021 gtggtgagct gaaactgctg ctgctgggtc cgggtgaatc tggcaaatcc accttcatta
     1081 aacagatgcg tatcatccac ggtgctggct actccgagga agaacgtaaa ggttttcgcc
     1141 cgctggttta tcagaatatc tttgtaagca tgcgtgcgat gatcgaagct atggaacgtc
     1201 tgcagatccc gttctcccgt ccggaatcta aacaccacgc gtccctggtg atgagccagg
     1261 acccgtacaa agttactact ttcgagaaac gttatgccgc agctatgcaa tggctgtggc
     1321 gtgacgcagg catccgtgct tactatgaac gtcgtcgcga attccacctg ctggattctg
     1381 cggtctatta cctgagccat ctggaacgta tcaccgaaga aggctacgtt ccaaccgcac
     1441 aggacgtact gcgctctcgc atgccgacca cgggtatcaa cgaatactgc ttctccgtgc
     1501 agaaaaccaa cctgcgcatc gttgatgtcg gtggccagaa aagcgaacgc aaaaagtgga
     1561 ttcattgttt cgaaaacgtg atcgcactga tctacctggc gtctctgagc gaatatgatc
     1621 aatgcctgga ggaaaacaat caggagaacc gtatgaaaga aagcctggcg ctgttcggta
     1681 ccattctgga gctgccgtgg tttaaatcta cttccgttat tctgttcctg aacaagaccg
     1741 atatcctgga agaaaaaatc ccgacgtccc acctggccac ttatttcccg tccttccagg
     1801 gccctaaaca ggacgcggaa gcagccaagc gttttattct ggacatgtac actcgtatgt
     1861 acaccggttg tgtggacggc ccggaaggca gcaagaaagg tgcacgctct cgtcgcctgt
     1921 tctctcacta cacctgcgct accgatacgc agaacatccg caaagtgttt aaagacgttc
     1981 gtgattctgt actggcgcgc tacctggatg aaatcaacct gctggcggcc gcataataaa
     2041 ccgatacaat taaaggctcc ttttggagcc ttttttttgg agattttcaa cgtgaaaaaa
     2101 ttattattcg caattccttt agttgttcct ttctattctc actccgctga aactgttgaa
     2161 agttgtttag caaaacccca tacagaaaat tcatttacta acgtctggaa agacgacaaa
     2221 actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg cgttgtagtt
     2281 tgtactggtg acgaaactca gtgtctagct agagtggcgg tggctctggt tccggtgatt
     2341 ttgattatga aaagatggca aacgctaata agggggctat gaccgaaaat gccgatgaaa
     2401 acgcgctaca gtctgacgct aaaggcaaac ttgattctgt cgctactgat tacggtgctg
     2461 ctatcgatgg tttcattggt gacgtttccg gccttgctaa tggtaatggt gctactggtg
     2521 attttgctgg ctctaattcc caaatggctc aagtcggtga cggtgataat tcacctttaa
     2581 tgaataattt ccgtcaatat ttaccttccc tccctcaatc ggttgaatgt cgcccttttg
     2641 tctttggcgc tggtaaacca tatgaatttt ctattgattg tgacaaaata aacttattcc
     2701 gtggtgtctt tgcgtttctt ttatatgttg ccacctttat gtatgtattt tctacgtttg
     2761 ctaacatact gcgtaataag gagtcttaat catgccagtt cttttggcta gcgccgccct
     2821 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     2881 aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa
     2941 ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc
     3001 gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg
     3061 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     3121 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     3181 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     3241 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     3301 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     3361 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     3421 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     3481 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     3541 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     3601 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     3661 ctttaccgca ggatccgcga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3721 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3781 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3841 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3901 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3961 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4021 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4081 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgg
     4141 atcctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4201 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4261 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4321 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4381 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     4441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4681 acaggactat aaagatacca gggtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4801 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5041 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5341 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5401 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5461 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5521 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5581 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5641 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     5701 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5761 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5821 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5881 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5941 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     6001 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6061 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6361 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6421 cctttcgtct tcaagaattc ccgacaccat cgaatggtgc aaaacctttc gcggtatggc
     6481 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     6541 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     6601 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     6661 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     6721 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     6781 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     6841 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     6901 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     6961 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     7021 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     7081 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     7141 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     7201 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     7261 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     7321 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     7381 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     7441 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     7501 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     7561 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     7621 taagttagct cactcattag gcacaattct catgtttgac agcttatcat cgactgcacg
     7681 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     7741 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
     7801 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctg
//
