LOCUS       sequence-485883-        6669 bp    DNA     circular SYN 12-SEP-2025
DEFINITION  Expresses GRP78 fused to the photoactivatable Tag-red fluorescent 
            protein (PATagRFP-GRP78).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6669)
  AUTHORS   Tseng CC, Stanciauskas R, Zhang P, Woo D, Wu K, Kelly K, Gill PS, Yu
            M, Pinaud F, Lee AS
  TITLE     GRP78 regulates CD44v membrane homeostasis and cell spreading in 
            tamoxifen-resistant breast cancer.
  JOURNAL   Life Sci Alliance. 2019 Aug 15;2(4):e201900377. doi: 
            10.26508/lsa.201900377. Print 2019 Aug.
  PUBMED    31416894
REFERENCE   2  (bases 1 to 6669)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Life Sci 
            Alliance."; date: "2019-08-15"; volume: "2(4):e201900377. doi"; 
            pages: " 10.26508/lsa.201900377. Print 2019 Aug"
COMMENT     Sequence Label: PATagRFP-GRP78
FEATURES             Location/Qualifiers
     source          1..6669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             629..1327
                     /codon_start=1
                     /product="photoactivatable variant of the monomeric red 
                     fluorescent protein TagRFP (Subach et al., 2010)"
                     /label=PA-TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSSTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMKKKTLGWEPSTEKLKPADGGLEGRVDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDRRLEIIKEADKETYWEQHEVAVARYS
                     DLPSKLGHKLN"
     polyA_signal    3397..3518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3434..3453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3488..3507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3525..3980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3662..3683)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3874..3893
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4007..4111
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4113..4470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4113..4304
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4136..4156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4321..4456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4505..5299
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4505..5299
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4559..4578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5169..5188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5487..5506)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5531..5578
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5907..6495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6396..6415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgaag ctctccctgg tggccgcgat
      601 gctgctgctg ctcagcgcgg cgcgggccat gagcgagctg attaaggaga acatgcacat
      661 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
      721 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
      781 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcagca ccttcatcaa
      841 ccacacccag ggcatccccg acttctggaa gcagtccttc cctgagggct tcacatggga
      901 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
      961 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
     1021 tgtgatgaag aagaaaacac tcggctggga accgagcacc gagaaactga aacccgctga
     1081 cggcggcctg gaaggcaggg tcgacatggc cctgaagctc gtgggcgggg gccacctgat
     1141 ctgcaacttc aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg
     1201 cgtctactat gtggaccgca gactggaaat catcaaggag gccgacaaag agacctactg
     1261 ggagcagcac gaggtggctg tggccagata ctccgacctc cctagcaaac tggggcacaa
     1321 gcttaatccg cggtccggag cggccgctgc aggaggcagc caaaaagagg aggaggacaa
     1381 gaaggaggac gtgggcacgg tggtcggcat cgacctgggg accacctact cctgcgtcgg
     1441 cgtgttcaag aacggccgcg tggagatcat cgccaacgat cagggcaacc gcatcacgcc
     1501 gtcctatgtc gccttcactc ctgaagggga acgtctgatt ggcgatgccg ccaagaacca
     1561 gctcacctcc aaccccgaga acacggtctt tgacgccaag cggctcatcg gccgcacgtg
     1621 gaatgacccg tctgtgcagc aggacatcaa gttcttgccg ttcaaggtgg ttgaaaagaa
     1681 aactaaacca tacattcaag ttgatattgg aggtgggcaa acaaagacat ttgctcctga
     1741 agaaatttct gccatggttc tcactaaaat gaaagaaacc gctgaggctt atttgggaaa
     1801 gaaggttacc catgcagttg ttactgtacc agcctatttt aatgatgccc aacgccaagc
     1861 aaccaaagac gctggaacta ttgctggcct aaatgttatg aggatcatca acgagcctac
     1921 ggcagctgct attgcttatg gcctggataa gagggagggg gagaagaaca tcctggtgtt
     1981 tgacctgggt ggcggaacct tcgatgtgtc tcttctcacc attgacaatg gtgtcttcga
     2041 agttgtggcc actaatggag atactcatct gggtggagaa gactttgacc agcgtgtcat
     2101 ggaacacttc atcaaactgt acaaaaagaa gacgggcaaa gatgtcagga aagacaatag
     2161 agctgtgcag aaactccggc gcgaggtaga aaaggccaaa cgggccctgt cttctcagca
     2221 tcaagcaaga attgaaattg agtccttcta tgaaggagaa gacttttctg agaccctgac
     2281 tcgggccaaa tttgaagagc tcaacatgga tctgttccgg tctactatga agcccgtcca
     2341 gaaagtgttg gaagattctg atttgaagaa gtctgatatt gatgaaattg ttcttgttgg
     2401 tggctcgact cgaattccaa agattcagca actggttaaa gagttcttca atggcaagga
     2461 accatcccgt ggcataaacc cagatgaagc tgtagcgtat ggtgctgctg tccaggctgg
     2521 tgtgctctct ggtgatcaag atacaggtga cctggtactg cttgatgtat gtccccttac
     2581 acttggtatt gaaactgtgg gaggtgtcat gaccaaactg attccaagga acacagtggt
     2641 gcctaccaag aagtctcaga tcttttctac agcttctgat aatcaaccaa ctgttacaat
     2701 caaggtctat gaaggtgaaa gacccctgac aaaagacaat catcttctgg gtacatttga
     2761 tctgactgga attcctcctg ctcctcgtgg ggtcccacag attgaagtca cctttgagat
     2821 agatgtgaat ggtattcttc gagtgacagc tgaagacaag ggtacaggga acaaaaataa
     2881 gatcacaatc accaatgacc agaatcgcct gacacctgaa gaaatcgaaa ggatggttaa
     2941 tgatgctgag aagtttgctg aggaagacaa aaagctcaag gagcgcattg atactagaaa
     3001 tgagttggaa agctatgcct attctctaaa gaatcagatt ggagataaag aaaagctggg
     3061 aggtaaactt tcctctgaag ataaggagac catggaaaaa gctgtagaag aaaagattga
     3121 atggctggaa agccaccaag atgctgacat tgaagacttc aaagctaaga agaaggaact
     3181 ggaagaaatt gttcaaccaa ttatcagcaa actctatgga agtgcaggcc ctcccccaac
     3241 tggtgaagag gatacagcag aaaaagatga gttgtagggt acctgatcta gatcataatc
     3301 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3361 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3421 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3481 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     3541 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3601 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3661 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3721 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3781 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3841 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3901 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3961 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4021 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4081 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     4141 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4261 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4381 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4441 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4501 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4561 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4621 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4681 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4741 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4801 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4861 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4921 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4981 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     5041 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5101 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5161 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5221 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5281 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5341 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5401 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5461 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5521 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5581 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5641 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5701 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5761 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5821 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5881 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5941 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6001 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6061 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6121 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6181 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6241 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6301 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6361 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6421 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6481 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6541 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6601 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     6661 ggagttccg
//
