LOCUS       sequence-482367-        6124 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expresses GRP78 N terminus fused to the GST Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6124)
  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 6124)
  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-N Terminus (pGEX4T1)
FEATURES             Location/Qualifiers
     source          1..6124
                     /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(2172..2194)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2339..2357)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2425..2529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2530..2598,2599..3390)
                     /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(2748..2767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3561..4149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4050..4069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4303..4320
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4393..4470
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4471..5553
                     /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(4490..4509)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5602..5607,5608..5625,5626..5632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5640..5656
                     /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     5664..5680
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5664..5680
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5676..6124,1..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(5696..5713)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5696..5712)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5705..5727)
                     /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 ggttagctcg agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat
     2161 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2221 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2281 gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc
     2341 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2401 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2461 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2521 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2581 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2641 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2701 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2761 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2821 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2881 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2941 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3001 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3061 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3121 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3181 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3241 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3301 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3361 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3421 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3481 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3541 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3601 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3661 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtyct tctagtgtag
     3721 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3781 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3841 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3901 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3961 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4021 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4081 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4141 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4201 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4261 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4321 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4381 cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
     4441 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     4501 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     4561 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     4621 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     4681 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     4741 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     4801 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     4861 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     4921 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     4981 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     5041 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     5101 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     5161 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     5221 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     5281 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     5341 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     5401 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     5461 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5521 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     5581 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5641 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg
     5701 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5761 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5821 cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag
     5881 cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct
     5941 caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta
     6001 cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     6061 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa
     6121 ttag
//
