LOCUS       sequence-215059-        7854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of GNA13_G-alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7854)
  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 7854)
  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_GNA13_G-alpha
FEATURES             Location/Qualifiers
     source          1..7854
                     /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      2049..2096
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3689..4144)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3847)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4038..4057
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4186..4208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4294..4434
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4349..4368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4449..4466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4620..5207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5378..6169,6170..6238))
                     /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     6001..6020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6239..6343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6411..6429
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6452..6529
                     /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             6530..7612
                     /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(6549..6568)
                     /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 cggccgccga tttcctgccg tctcgtagcg tcctgagtgt ctgctttccg ggttgcctgc
      961 tgaccagtgg cgaagctgaa caacagcgta agtccaaaga aattgataag tgcctgtcac
     1021 gcgaaaaaac ctatgtgaag cgtctggtta aaatcctgct gctgggcgcc ggtgaatccg
     1081 gcaagtcaac gtttctgaaa cagatgcgca ttatccatgg tcaagatttt gaccagcgcg
     1141 cccgtgaaga attccgtccg accatttaca gcaacgttat caagggtatg cgcgtgctgg
     1201 ttgatgcacg tgaaaaactg catattccgt ggggcgacaa ctcgaatcag caacacggtg
     1261 ataaaatgat gagctttgac acccgcgctc cgatggcggc acagggtatg gtcgaaacgc
     1321 gcgtgttcct gcagtatctg ccggcaattc gtgctctgtg ggcggattct ggcatccaaa
     1381 acgcgtacga ccgtcgccgt gaatttcagc tgggtgaaag tgtgaaatat ttcctggata
     1441 atctggacaa gctgggcgaa ccggattaca ttccgtccca gcaagacatc ctgctggcac
     1501 gccgtccgac caagggtatc cacgaatacg atttcgaaat caagaacgtt ccgttcaaga
     1561 tggtcgacgt gggcggtcag cgctctgaac gcaaacgttg gtttgaatgt ttcgattcgg
     1621 tcaccagcat tctgtttctg gttagctcta gtgaattcga tcaggtgctg atggaagacc
     1681 gcctgaccaa ccgtctgacg gaaagcctga acatcttcga aaccatcgtt aacaaccgtg
     1741 tcttttctaa cgttagtatt atcctgttcc tgaataagac ggatctgctg gaagaaaaag
     1801 ttcagattgt cagtatcaag gattattttc tggaattcga aggcgatccg cattgcctgc
     1861 gcgacgtgca gaaatttctg gttgaatgtt tccgtaacaa acgccgtgat cagcaacaga
     1921 agccgctgta ccatcacttt accacggcga ttaacaccga aaatatccgc ctggttttcc
     1981 gtgatgtcaa agacacgatc ctgcacgata atctgaagca actgatgctg caggcggccg
     2041 cataataaac cgatacaatt aaaggctcct tttggagcct tttttttgga gattttcaac
     2101 gtgaaaaaat tattattcgc aattccttta gttgttcctt tctattctca ctccgctgaa
     2161 actgttgaaa gttgtttagc aaaaccccat acagaaaatt catttactaa cgtctggaaa
     2221 gacgacaaaa ctttagatcg ttacgctaac tatgagggct gtctgtggaa tgctacaggc
     2281 gttgtagttt gtactggtga cgaaactcag tgtctagcta gagtggcggt ggctctggtt
     2341 ccggtgattt tgattatgaa aagatggcaa acgctaataa gggggctatg accgaaaatg
     2401 ccgatgaaaa cgcgctacag tctgacgcta aaggcaaact tgattctgtc gctactgatt
     2461 acggtgctgc tatcgatggt ttcattggtg acgtttccgg ccttgctaat ggtaatggtg
     2521 ctactggtga ttttgctggc tctaattccc aaatggctca agtcggtgac ggtgataatt
     2581 cacctttaat gaataatttc cgtcaatatt taccttccct ccctcaatcg gttgaatgtc
     2641 gcccttttgt ctttggcgct ggtaaaccat atgaattttc tattgattgt gacaaaataa
     2701 acttattccg tggtgtcttt gcgtttcttt tatatgttgc cacctttatg tatgtatttt
     2761 ctacgtttgc taacatactg cgtaataagg agtcttaatc atgccagttc ttttggctag
     2821 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     2881 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga
     2941 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     3001 atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg
     3061 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
     3121 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
     3181 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
     3241 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
     3301 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga
     3361 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc
     3421 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc
     3481 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag
     3541 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
     3601 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
     3661 gctgatgagc tttaccgcag gatccgcgaa attgtaagcg ttaatatttt gttaaaattc
     3721 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3781 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3841 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3901 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3961 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4021 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4081 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4141 gcgtcgcgga tcctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4201 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4261 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     4321 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4381 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     4441 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4501 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4561 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4621 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4681 cgaaacccga caggactata aagataccag ggtttccccc tggaagctcc ctcgtgcgct
     4741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5041 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5221 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5281 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     5701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     6001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6361 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6421 tcacgaggcc ctttcgtctt caagaattcc cgacaccatc gaatggtgca aaacctttcg
     6481 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     6541 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     6601 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     6661 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     6721 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     6781 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     6841 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     6901 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     6961 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     7021 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     7081 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     7141 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     7201 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     7261 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     7321 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     7381 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     7441 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     7501 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7561 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7621 caattaatgt aagttagctc actcattagg cacaattctc atgtttgaca gcttatcatc
     7681 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
     7741 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     7801 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctg
//
