LOCUS       sequence-482354-        6829 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expresses GRP78 with the last 11 amino acids at the C terminus 
            deleted fused to the GST Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  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 6829)
  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-Delta C11(pGEX4T1)
FEATURES             Location/Qualifiers
     source          1..6829
                     /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(2877..2899)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3044..3062)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3130..3234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3235..3303,3304..4095)
                     /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(3453..3472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4266..4854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4755..4774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5008..5025
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5098..5175
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5176..6258
                     /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(5195..5214)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6307..6312,6313..6330,6331..6337)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6345..6361
                     /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     6369..6385
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6369..6385
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6381..6829,1..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(6401..6418)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6401..6417)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6410..6432)
                     /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 tccgaggagg aggacaagaa ggaggacgtg
      961 ggcacggtgg tcggcatcga cctggggacc acctactcct gcgtcggcgt gttcaagaac
     1021 ggccgcgtgg agatcatcgc caacgatcag ggcaaccgca tcacgccgtc ctatgtcgcc
     1081 ttcactcctg aaggggaacg tctgattggc gatgccgcca agaaccagct cacctccaac
     1141 cccgagaaca cggtctttga cgccaagcgg ctcatcggcc gcacgtggaa tgacccgtct
     1201 gtgcagcagg acatcaagtt cttgccgttc aaggtggttg aaaagaaaac taaaccatac
     1261 attcaagttg atattggagg tgggcaaaca aagacatttg ctcctgaaga aatttctgcc
     1321 atggttctca ctaaaatgaa agaaaccgct gaggcttatt tgggaaagaa ggttacccat
     1381 gcagttgtta ctgtaccagc ctattttaat gatgcccaac gccaagcaac caaagacgct
     1441 ggaactattg ctggcctaaa tgttatgagg atcatcaacg agcctacggc agctgctatt
     1501 gcttatggcc tggataagag ggagggggag aagaacatcc tggtgtttga cctgggtggc
     1561 ggaaccttcg atgtgtctct tctcaccatt gacaatggtg tcttcgaagt tgtggccact
     1621 aatggagata ctcatctggg tggagaagac tttgaccagc gtgtcatgga acacttcatc
     1681 aaactgtaca aaaagaagac gggcaaagat gtcaggaaag acaatagagc tgtgcagaaa
     1741 ctccggcgcg aggtagaaaa ggccaaacgg gccctgtctt ctcagcatca agcaagaatt
     1801 gaaattgagt ccttctatga aggagaagac ttttctgaga ccctgactcg ggccaaattt
     1861 gaagagctca acatggatct gttccggtct actatgaagc ccgtccagaa agtgttggaa
     1921 gattctgatt tgaagaagtc tgatattgat gaaattgttc ttgttggtgg ctcgactcga
     1981 attccaaaga ttcagcaact ggttaaagag ttcttcaatg gcaaggaacc atcccgtggc
     2041 ataaacccag atgaagctgt agcgtatggt gctgctgtcc aggctggtgt gctctctggt
     2101 gatcaagata caggtgacct ggtactgctt gatgtatgtc cccttacact tggtattgaa
     2161 actgtgggag gtgtcatgac caaactgatt ccaaggaaca cagtggtgcc taccaagaag
     2221 tctcagatct tttctacagc ttctgataat caaccaactg ttacaatcaa ggtctatgaa
     2281 ggtgaaagac ccctgacaaa agacaatcat cttctgggta catttgatct gactggaatt
     2341 cctcctgctc ctcgtggggt cccacagatt gaagtcacct ttgagataga tgtgaatggt
     2401 attcttcgag tgacagctga agacaagggt acagggaaca aaaataagat cacaatcacc
     2461 aatgaccaga atcgcctgac acctgaagaa atcgaaagga tggttaatga tgctgagaag
     2521 tttgctgagg aagacaaaaa gctcaaggag cgcattgata ctagaaatga gttggaaagc
     2581 tatgcctatt ctctaaagaa tcagattgga gataaagaaa agctgggagg taaactttcc
     2641 tctgaagata aggagaccat ggaaaaagct gtagaagaaa agattgaatg gctggaaagc
     2701 caccaagatg ctgacattga agacttcaaa gctaagaaga aggaactgga agaaattgtt
     2761 caaccaatta tcagcaaact ctatggaagt gcaggccctc ccccaactta gctcgagcgg
     2821 ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     2881 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     2941 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     3001 tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta
     3061 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3121 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3181 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3241 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3301 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3361 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3421 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     3481 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3541 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3601 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3661 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3721 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     3781 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3841 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3901 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3961 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4021 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4081 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4141 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4201 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4261 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4321 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4381 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4441 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4501 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4561 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4621 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4681 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4741 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4801 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4861 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4921 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4981 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5041 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac
     5101 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     5161 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     5221 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     5281 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     5341 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     5401 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     5461 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     5521 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     5581 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     5641 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     5701 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     5761 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     5821 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     5881 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     5941 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     6001 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     6061 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     6121 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     6181 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6241 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
     6301 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6361 atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac
     6421 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6481 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6541 gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct
     6601 ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc
     6661 acggttacga tgcgcccatc tacaccaacg tgacctatcc cattacggtc aatccgccgt
     6721 ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct
     6781 ggctacagga aggccagacg cgaattattt ttgatggcgt tggaattag
//
