LOCUS       sequence-472912-        7434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GRP78 Full Length without UTRs fused to the FLAG Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7434)
  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 7434)
  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.neo.2019.05.008"; journalName: "Neoplasia"; date: 
            "2019-08"; volume: "21"; issue: "8"; pages: "837-848"
COMMENT     Sequence Label: FLAG-GRP78-No UTRs Wild-Type (pcDNA3.1)
FEATURES             Location/Qualifiers
     source          1..7434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             967..990
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2987..3005)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2988..3005)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3025..3042)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3031..3255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3301..3729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3388..3407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3598..3619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3738..3758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3743..4072
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3923..4058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3985..4004
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4139..4933
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4193..4212)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4803..4822
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5107..5228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5144..5163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5198..5217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5277..5293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5277..5293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5290..5312)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5301..5317
                     /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)."
     promoter        complement(join(5325..5331,5332..5349,5350..5355))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5370..5391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5508..5525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5679..6267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5759..5778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6438..7229,7230..7298))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7061..7080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7299..7403)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccatgaagct ctccctggtg gccgcgatgc tgctgctgct cagcgcggcg
      961 cgggccgact acaaagatga cgatgacaag gaggaggagg acaagaagga ggacgtgggc
     1021 acggtggtcg gcatcgacct ggggaccacc tactcctgcg tcggcgtgtt caagaacggc
     1081 cgcgtggaga tcatcgccaa cgatcagggc aaccgcatca cgccgtccta tgtcgccttc
     1141 actcctgaag gggaacgtct gattggcgat gccgccaaga accagctcac ctccaacccc
     1201 gagaacacgg tctttgacgc caagcggctc atcggccgca cgtggaatga cccgtctgtg
     1261 cagcaggaca tcaagttctt gccgttcaag gtggttgaaa agaaaactaa accatacatt
     1321 caagttgata ttggaggtgg gcaaacaaag acatttgctc ctgaagaaat ttctgccatg
     1381 gttctcacta aaatgaaaga aaccgctgag gcttatttgg gaaagaaggt tacccatgca
     1441 gttgttactg taccagccta ttttaatgat gcccaacgcc aagcaaccaa agacgctgga
     1501 actattgctg gcctaaatgt tatgaggatc atcaacgagc ctacggcagc tgctattgct
     1561 tatggcctgg ataagaggga gggggagaag aacatcctgg tgtttgacct gggtggcgga
     1621 accttcgatg tgtctcttct caccattgac aatggtgtct tcgaagttgt ggccactaat
     1681 ggagatactc atctgggtgg agaagacttt gaccagcgtg tcatggaaca cttcatcaaa
     1741 ctgtacaaaa agaagacggg caaagatgtc aggaaagaca atagagctgt gcagaaactc
     1801 cggcgcgagg tagaaaaggc caaacgggcc ctgtcttctc agcatcaagc aagaattgaa
     1861 attgagtcct tctatgaagg agaagacttt tctgagaccc tgactcgggc caaatttgaa
     1921 gagctcaaca tggatctgtt ccggtctact atgaagcccg tccagaaagt gttggaagat
     1981 tctgatttga agaagtctga tattgatgaa attgttcttg ttggtggctc gactcgaatt
     2041 ccaaagattc agcaactggt taaagagttc ttcaatggca aggaaccatc ccgtggcata
     2101 aacccagatg aagctgtagc gtatggtgct gctgtccagg ctggtgtgct ctctggtgat
     2161 caagatacag gtgacctggt actgcttgat gtatgtcccc ttacacttgg tattgaaact
     2221 gtgggaggtg tcatgaccaa actgattcca aggaacacag tggtgcctac caagaagtct
     2281 cagatctttt ctacagcttc tgataatcaa ccaactgtta caatcaaggt ctatgaaggt
     2341 gaaagacccc tgacaaaaga caatcatctt ctgggtacat ttgatctgac tggaattcct
     2401 cctgctcctc gtggggtccc acagattgaa gtcacctttg agatagatgt gaatggtatt
     2461 cttcgagtga cagctgaaga caagggtaca gggaacaaaa ataagatcac aatcaccaat
     2521 gaccagaatc gcctgacacc tgaagaaatc gaaaggatgg ttaatgatgc tgagaagttt
     2581 gctgaggaag acaaaaagct caaggagcgc attgatacta gaaatgagtt ggaaagctat
     2641 gcctattctc taaagaatca gattggagat aaagaaaagc tgggaggtaa actttcctct
     2701 gaagataagg agaccatgga aaaagctgta gaagaaaaga ttgaatggct ggaaagccac
     2761 caagatgctg acattgaaga cttcaaagct aagaagaagg aactggaaga aattgttcaa
     2821 ccaattatca gcaaactcta tggaagtgca ggccctcccc caactggtga agaggataca
     2881 gcagaaaaag atgagttgta gacactgatc tgctagtgct gtaatattgt aaatactgga
     2941 ctcaggaact tttgttaggc tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     3001 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3061 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3121 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3181 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3241 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3361 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3481 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3601 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3661 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3721 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3901 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3961 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4021 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4081 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4141 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4201 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4261 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4321 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4381 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4441 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4501 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4561 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4621 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4681 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4741 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4801 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4861 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4921 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4981 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5041 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5101 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5161 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5221 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5281 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5341 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5401 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5461 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5521 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5581 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5641 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5701 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5761 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5821 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5881 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5941 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6001 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6061 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6121 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6181 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6241 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6301 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6361 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6421 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6481 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6541 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6601 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6661 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6721 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6781 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6841 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6901 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6961 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7021 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7081 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7141 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7201 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7261 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7321 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7381 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
