LOCUS       sequence-288895-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsSmg6_1-576.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6706)
  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"
COMMENT     Sequence Label: pGEX-6P-1-HsSmg6_1-576_H
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2756..2778)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2923..2941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3009..3113
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3114..3182,3183..3974)
                     /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(3332..3351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4145..4733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4887..4904
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4977..5054
                     /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             5055..6137
                     /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(5074..5093)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6150..6171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6186..6191,6192..6209,6210..6216)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6224..6240
                     /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     6248..6264
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6248..6264
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6260..6706,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6280..6297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6280..6296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6289..6311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ccggaattga
      961 tggcggaagg gctggagcgt gtgcggatct ccgcgtcgga gctgcgcggg atcctggcta
     1021 ctctggcccc gcaggccggg agcagagaaa acatgaagga attaaaggag gccaggccgc
     1081 gcaaagataa caggcgtcca gatctggaaa tctataagcc tggcctttct cggctaagga
     1141 acaagcccaa aatcaaggaa ccccctggga gtgaggaatt caaagatgaa attgttaatg
     1201 accgagattg ctctgctgtt gaaaatggta cacagcccgt taaagatgtc tgcaaggaac
     1261 tgaacaacca agagcagaat ggtcctatag acccagaaaa taatcgggga caagaatcct
     1321 ttcctaggac tgctggacaa gaggatcgta gtctaaaaat tatcaaaaga acaaagaaac
     1381 ccgacctgca gatctatcag cctggacgac gtttgcagac tgttagcaaa gaatccgcca
     1441 gtcgggtgga ggaggaagaa gtcctcaacc aggtagaaca actgagagta gaggaagatg
     1501 agtgtagggg aaatgttgcg aaggaggaag ttgcgaataa accagacagg gccgagatag
     1561 aaaagagccc aggtggtggg agagtagggg ctgcaaaagg agaaaaagga aagaggatgg
     1621 gaaaagggga gggggtgagg gaaacccacg acgacccggc ccgcgggagg ccgggctccg
     1681 caaagcgcta ctcccgctca gacaaacgaa ggaatcgcta ccgcacgcgc agcaccagct
     1741 cagctggcag caacaacagc gctgagggag ctggcctgac ggataatgga tgtcgccgcc
     1801 gccgacagga taggaccaag gagaggccac cactgaagaa gcaagtgtct gtgtcctcaa
     1861 ccgattcctt agacgaggac agaattgatg agcctgatgg attaggaccc aggagaagtt
     1921 cagaaaggaa gagacattta gaaagaaact ggtctggccg tggggagggt gagcagaaaa
     1981 ccagtgctaa agaatatcga ggcactcttc gtgtcacttt cgatgcagaa gccatgaaca
     2041 aagagtctcc catggtgagg tcagccaggg atgatatgga tagaggaaag cctgacaaag
     2101 gcttgagcag tgggggcaaa ggctctgaga agcaggagtc caaaaacccg aaacaagaac
     2161 ttcggggtcg tggtcgtggc attctgattt tgcctgccca taccacccta tctgtcaatt
     2221 cagcaggttc tccagagtcc gcgcctttgg gacctcggct tttgtttgga tctggtagta
     2281 agggatctcg gagttggggc cgtggaggca ccacacgccg attgtgggac ccaaacaatc
     2341 ctgatcagaa acctgctcta aagactcaga cgccccagct acatttcttg gacactgatg
     2401 atgaagtcag ccctacatct tggggtgact cacgccaggc tcaggcatct tactataagt
     2461 ttcaaaactc tgacaacccc tattattacc cccggacacc aggccctgcc tcccagtatc
     2521 cctatacggg ctataaccct ctacagtacc cagtgggccc tacgaatggt gtgtacccag
     2581 ggccttacta cccaggctac ccgactccgt caggacagta tgtgtgtagc cctctaccta
     2641 ccagcaccat gagtcccgag gaggtagagc agcacatgag gaacctgtga ctcgagcggc
     2701 cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     2761 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2821 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     2881 cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat
     2941 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3001 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3061 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3121 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3181 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3241 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3301 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     3361 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3421 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3481 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3541 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3601 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     3661 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3721 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3781 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3841 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3901 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3961 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4021 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4081 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4141 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4201 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4261 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4321 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4381 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4441 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4501 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4561 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4621 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4681 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4741 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4801 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4861 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4921 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca
     4981 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     5041 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     5101 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     5161 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     5221 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     5281 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     5341 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     5401 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     5461 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     5521 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     5581 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     5641 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5701 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     5761 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     5821 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     5881 ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat tttcgcctgc
     5941 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     6001 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     6061 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6121 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6181 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6241 tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg
     6301 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     6361 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     6421 cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg
     6481 gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca
     6541 cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca atccgccgtt
     6601 tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     6661 gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaatt
//
