LOCUS       sequence-175728-        5844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human progesterone receptor ligand binding 
            domain amino acid residues 637-933 with GST tag in pGEX-3X-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5844)
  TITLE     Wilson Lab plasmids
REFERENCE   2  (bases 1 to 5844)
  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: GST-PR-LBD
FEATURES             Location/Qualifiers
     source          1..5844
                     /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)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             739..750
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1716..1738)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1879..1897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1965..2069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2070..2138,2139..2930)
                     /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(2288..2307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3101..3689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3590..3609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3843..3860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3933..4010
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4011..5093
                     /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(4030..4049)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5142..5147,5148..5165,5166..5172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5180..5196
                     /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     5204..5220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5204..5220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5216..5743
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5236..5253)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5236..5252)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5245..5267)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctgat cgaaggtcgt gggatccgaa aatttaaaaa gttcaataaa
      781 gtcagagttg tgagagcact ggatgctgtt gctctcccac agccattggg cgttccaaat
      841 gaaagccaag ccctaagcca gagattcact ttttcaccag gtcaagacat acagttgatt
      901 ccaccactga tcaacctgtt aatgagcatt gaaccagatg tgatctatgc aggacatgac
      961 aacacaaaac ctgacacctc cagttctttg ctgacaagtc ttaatcaact aggcgagagg
     1021 caacttcttt cagtagtcaa gtggtctaaa tcattgccag gttttcgaaa cttacatatt
     1081 gatgaccaga taactctcat tcagtattct tggatgagct taatggtgtt tggtctagga
     1141 tggagatcct acaaacatgt cagtgggcag atgctgtatt ttgcacctga tctaatacta
     1201 aatgaacagc ggatgaaaga atcatcattc tattcattat gccttaccat gtggcagatc
     1261 ccacaggagt ttgtcaagct tcaagttagc caagaagagt tcctctgtat gaaagtattg
     1321 ttacttctta atacaattcc tttggaaggg ctacgaagtc aaacccagtt tgaggagatg
     1381 aggtcaagct acattagaga gctcatcaag gcaattggtt tgaggcaaaa aggagttgtg
     1441 tcgagctcac agcgtttcta tcaacttaca aaacttcttg ataacttgca tgatcttgtc
     1501 aaacagcttc atctgtactg cttgaataca tttatccagt cccgggcact gagtgttgaa
     1561 tttccagaaa tgatgtctga agttattgct gcacaattac ccaagatatt ggcagggatg
     1621 gtgaaacccc ttctctttca taaaaagtga atgtctagaa ttcatcgtga ctgactgacg
     1681 atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     1741 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     1801 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagta
     1861 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1921 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1981 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2041 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2101 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2161 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2221 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2281 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     2341 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2401 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2461 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2521 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2581 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     2641 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2701 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2761 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2821 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2881 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2941 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3001 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3061 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3121 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3181 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3241 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3301 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3361 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3421 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3481 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3541 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3601 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3661 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3721 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3781 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3841 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3901 atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc
     3961 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     4021 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     4081 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     4141 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     4201 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     4261 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     4321 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     4381 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     4441 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     4501 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     4561 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     4621 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     4681 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     4741 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     4801 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     4861 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     4921 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     4981 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5041 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5101 gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     5161 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     5221 catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5281 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5341 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc
     5401 ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc
     5461 cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac
     5521 caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg
     5581 ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat
     5641 tatttttgat ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt
     5701 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt
     5761 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     5821 tggcaaatat tctgaaatga gctg
//
