LOCUS       sequence-482368-        6865 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expresses GRP78 Full Length fused to the GST Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6865)
  AUTHORS   Tseng CC, Zhang P, Lee AS
  TITLE     The COOH-Terminal Proline-Rich Region of GRP78 Is a Key Regulator of
            Its Cell Surface Expression and Viability of Tamoxifen-Resistant 
            Breast Cancer Cells.
  JOURNAL   Neoplasia. 2019 Aug;21(8):837-848. doi: 10.1016/j.neo.2019.05.008. 
            Epub 2019 Jul 12.
  PUBMED    31306849
REFERENCE   2  (bases 1 to 6865)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neo.2019.05.008"; journalName: "Neoplasia"; date: 
            "2019-08"; volume: "21"; issue: "8"; pages: "837-848"
COMMENT     Sequence Label: GST-GRP78-Full Length (pGEX4T1)
FEATURES             Location/Qualifiers
     source          1..6865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(181..186,187..202,203..209)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    217..233
                     /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             256..909
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     867..889
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             916..933
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2913..2935)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3080..3098)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3166..3270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3271..3339,3340..4131)
                     /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(3489..3508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4302..4890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4791..4810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5044..5061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5134..5211
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5212..6294
                     /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(5231..5250)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6343..6348,6349..6366,6367..6373)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6381..6397
                     /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     6405..6421
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6405..6421
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6417..6865,1..79)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6437..6454)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6437..6453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6446..6468)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
       61 atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
      121 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
      181 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
      241 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      301 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      361 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      421 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      481 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      541 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      601 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      661 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      721 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      781 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      841 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      901 cctccaaaat cggatctggt tccgcgtgga tccatggagg aggaggacaa gaaggaggac
      961 gtgggcacgg tggtcggcat cgacctgggg accacctact cctgcgtcgg cgtgttcaag
     1021 aacggccgcg tggagatcat cgccaacgat cagggcaacc gcatcacgcc gtcctatgtc
     1081 gccttcactc ctgaagggga acgtctgatt ggcgatgccg ccaagaacca gctcacctcc
     1141 aaccccgaga acacggtctt tgacgccaag cggctcatcg gccgcacgtg gaatgacccg
     1201 tctgtgcagc aggacatcaa gttcttgccg ttcaaggtgg ttgaaaagaa aactaaacca
     1261 tacattcaag ttgatattgg aggtgggcaa acaaagacat ttgctcctga agaaatttct
     1321 gccatggttc tcactaaaat gaaagaaacc gctgaggctt atttgggaaa gaaggttacc
     1381 catgcagttg ttactgtacc agcctatttt aatgatgccc aacgccaagc aaccaaagac
     1441 gctggaacta ttgctggcct aaatgttatg aggatcatca acgagcctac ggcagctgct
     1501 attgcttatg gcctggataa gagggagggg gagaagaaca tcctggtgtt tgacctgggt
     1561 ggcggaacct tcgatgtgtc tcttctcacc attgacaatg gtgtcttcga agttgtggcc
     1621 actaatggag atactcatct gggtggagaa gactttgacc agcgtgtcat ggaacacttc
     1681 atcaaactgt acaaaaagaa gacgggcaaa gatgtcagga aagacaatag agctgtgcag
     1741 aaactccggc gcgaggtaga aaaggccaaa cgggccctgt cttctcagca tcaagcaaga
     1801 attgaaattg agtccttcta tgaaggagaa gacttttctg agaccctgac tcgggccaaa
     1861 tttgaagagc tcaacatgga tctgttccgg tctactatga agcccgtcca gaaagtgttg
     1921 gaagattctg atttgaagaa gtctgatatt gatgaaattg ttcttgttgg tggctcgact
     1981 cgaattccaa agattcagca actggttaaa gagttcttca atggcaagga accatcccgt
     2041 ggcataaacc cagatgaagc tgtagcgtat ggtgctgctg tccaggctgg tgtgctctct
     2101 ggtgatcaag atacaggtga cctggtactg cttgatgtat gtccccttac acttggtatt
     2161 gaaactgtgg gaggtgtcat gaccaaactg attccaagga acacagtggt gcctaccaag
     2221 aagtctcaga tcttttctac agcttctgat aatcaaccaa ctgttacaat caaggtctat
     2281 gaaggtgaaa gacccctgac aaaagacaat catcttctgg gtacatttga tctgactgga
     2341 attcctcctg ctcctcgtgg ggtcccacag attgaagtca cctttgagat agatgtgaat
     2401 ggtattcttc gagtgacagc tgaagacaag ggtacaggga acaaaaataa gatcacaatc
     2461 accaatgacc agaatcgcct gacacctgaa gaaatcgaaa ggatggttaa tgatgctgag
     2521 aagtttgctg aggaagacaa aaagctcaag gagcgcattg atactagaaa tgagttggaa
     2581 agctatgcct attctctaaa gaatcagatt ggagataaag aaaagctggg aggtaaactt
     2641 tcctctgaag ataaggagac catggaaaaa gctgtagaag aaaagattga atggctggaa
     2701 agccaccaag atgctgacat tgaagacttc aaagctaaga agaaggaact ggaagaaatt
     2761 gttcaaccaa ttatcagcaa actctatgga agtgcaggcc ctcccccaac tggtgaagag
     2821 gatacagcag aaaaagatga gttgtagctc gagcggccgc atcgtgactg actgacgatc
     2881 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2941 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3001 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3061 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3121 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3241 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3301 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3361 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3421 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3481 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     3541 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3601 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3661 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3721 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3781 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     3841 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3901 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3961 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4021 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4441 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4921 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4981 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5041 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     5101 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
     5161 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     5221 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     5281 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     5341 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     5401 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     5461 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     5521 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     5581 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     5641 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     5701 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     5761 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     5821 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     5881 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     5941 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     6001 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     6061 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     6121 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     6181 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6241 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6301 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6361 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6421 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     6481 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     6541 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     6601 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     6661 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     6721 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     6781 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     6841 ttatttttga tggcgttgga attag
//
