LOCUS       sequence-272028-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of pGEX:PGRMC1 to perform a GST-pulldown assay 
            and identify new proteins that interact with GST-PGRMC1 in the human
            endometrial stroll cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  AUTHORS   Salsano S, Gonzalez-Martin R, Quinonero A, Lopez-Martin S, 
            Gomez-Escribano AP, Perez-Deben S, Yanez-Mo M, Dominguez F
  TITLE     Novel nonclassic progesterone receptor PGRMC1 pulldown-precipitated 
            proteins reveal a key role during human decidualization.
  JOURNAL   Fertil Steril. 2020 May;113(5):1050-1066.e7. doi: 
            10.1016/j.fertnstert.2020.01.008.
  PUBMED    32386616
REFERENCE   2  (bases 1 to 5566)
  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.fertnstert.2020.01"; journalName: "Fertil Steril"; date: 
            "2020-05"; volume: "113"; issue: "5"; pages: "1050-1066.e7"
COMMENT     Sequence Label: pGEX:PGRMC1
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(88..93,94..109,110..116)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    124..140
                     /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             163..816
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     774..796
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             823..846
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1521..1543)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1688..1706)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1774..1878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1879..1947,1948..2739)
                     /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(2097..2116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2910..3498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3399..3418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3652..3669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3742..3819
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3820..4902
                     /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(3839..3858)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4951..4956,4957..4974,4975..4981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4989..5005
                     /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     5013..5029
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5013..5029
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5025..5552
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5045..5062)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5045..5061)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5054..5076)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg
       61 ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg
      121 gaattgtgag cggataacaa tttcacacag gaaacagtat tcatgtcccc tatactaggt
      181 tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa
      241 aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt
      301 gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca
      361 cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca
      421 aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt
      481 tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta
      541 cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat
      601 catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac
      661 ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc
      721 ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg
      781 caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag
      841 gggcccctgg gatccatggc tgccgaggat gtggtggcga ctggcgccga cccaagcgat
      901 ctggagagcg gcgggctgct gcatgagatt ttcacgtcgc cgctcaacct gctgctgctt
      961 ggcctctgca tcttcctgct ctacaagatc gtgcgcgggg accagccggc ggccagcggc
     1021 gacagcgacg acgacgagcc gccccctctg ccccgcctca agcggcgcga cttcaccccc
     1081 gccgagctgc ggcgcttcga cggcgtccag gacccgcgca tactcatggc catcaacggc
     1141 aaggtgttcg atgtgaccaa aggccgcaaa ttctacgggc ccgaggggcc gtatggggtc
     1201 tttgctggaa gagatgcatc caggggcctt gccacatttt gcctggataa ggaagcactg
     1261 aaggatgagt acgatgacct ttctgacctc actgctgccc agcaggagac tctgagtgac
     1321 tgggagtctc agttcacttt caagtatcat cacgtgggca aactgctgaa ggagggggag
     1381 gagcccactg tgtactcaga tgaggaagaa ccaaaagatg agagtgcccg gaaaaatgat
     1441 gaattcccgg gtcgactcga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt
     1501 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     1561 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1621 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga
     1681 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1741 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1801 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1861 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1921 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1981 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2041 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2101 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2161 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2221 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2281 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2341 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2401 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     2461 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2521 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2581 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2641 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2701 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2761 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2821 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2881 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2941 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3001 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3061 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3121 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3181 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3241 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3301 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3361 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3421 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3481 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3541 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3601 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3661 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3721 atttcacacc gcataaattc cgacaccatc gaatggcgca aaacctttcg cggtatggca
     3781 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     3841 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     3901 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     3961 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     4021 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     4081 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     4141 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     4201 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     4261 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     4321 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     4381 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     4441 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     4501 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     4561 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     4621 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     4681 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     4741 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     4801 accacgctag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4861 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4921 aagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4981 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg
     5041 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     5101 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     5161 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
     5221 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg
     5281 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct
     5341 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc
     5401 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg
     5461 gcgttggaat tagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat
     5521 cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataatt
//
