LOCUS       sequence-482359-        6814 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expresses GRP78 with the last 17 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 6814)
  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 6814)
  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 C17 (pGEX4T1)
FEATURES             Location/Qualifiers
     source          1..6814
                     /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(2862..2884)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3029..3047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3115..3219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3220..3288,3289..4080)
                     /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(3438..3457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4251..4839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4740..4759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4993..5010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5083..5160
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5161..6243
                     /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(5180..5199)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6292..6297,6298..6315,6316..6322)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6330..6346
                     /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     6354..6370
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6354..6370
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6366..6814,1..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(6386..6403)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6386..6402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6395..6417)
                     /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 agttagctcg agcggccgca tcgtgactga
     2821 ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2881 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2941 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     3001 ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3061 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3121 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3181 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3241 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3301 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3361 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3421 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     3481 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3541 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3601 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3661 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3721 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     3781 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3841 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3901 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3961 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4021 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4081 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4141 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     5041 ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc
     5101 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     5161 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     5221 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     5281 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     5341 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     5401 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     5461 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     5521 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     5581 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     5641 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     5701 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     5761 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     5821 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     5881 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     5941 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     6001 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     6061 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     6121 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     6181 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6241 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
     6301 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6361 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     6421 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     6481 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6541 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
     6601 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
     6661 ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
     6721 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
     6781 agacgcgaat tatttttgat ggcgttggaa ttag
//
