LOCUS       sequence-314153-        6627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hamster BiP/GRP78/HSPA5 fused to mCherry. Useful for studying BiP 
            mobility and trafficking in live cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6627)
  AUTHORS   Lai CW, Aronson DE, Snapp EL
  TITLE     BiP availability distinguishes states of homeostasis and stress in 
            the endoplasmic reticulum of living cells.
  JOURNAL   Mol Biol Cell. 2010 Jun 15;21(12):1909-21. doi: 
            10.1091/mbc.E09-12-1066. Epub 2010 Apr 21.
  PUBMED    20410136
REFERENCE   2  (bases 1 to 6627)
  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: 
            "10.1091/mbc.E09-12-1066"; journalName: "Mol Biol Cell"; date: 
            "2010-06-15- 15"; volume: "21"; issue: "12"; pages: "1909-21"
COMMENT     Sequence Label: BiP-mCherry
FEATURES             Location/Qualifiers
     source          1..6627
                     /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"
     regulatory      2505..2514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2511..3218
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2656..2674)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2924..2943
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3355..3476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3392..3411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3446..3465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3483..3938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3620..3641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3832..3851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3965..4069
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4071..4428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4071..4262
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4094..4114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4279..4414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4463..5257
                     /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             4463..5257
                     /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(4517..4536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5127..5146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5445..5464)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5489..5536
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5865..6453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6354..6373
                     /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 gctagcacca
      541 tgaagttccc tatggtggcg gcggcgctgc tgctgctctg cgcggtgcgg gccgaggagg
      601 aggacaagaa ggaggatgtg ggcacggtgg tcggcatcga cctggggacc acctattcct
      661 gcgttggtgt gttcaagaac ggccgcgtgg agatcatagc caacgatcag ggcaaccgca
      721 tcacgccgtc gtatgtggcc ttcactcctg aaggcgagcg tctgattggc gatgcggcca
      781 agaaccagct cacctccaat cccgagaaca cggtcttcga cgccaagcgc ctcatcggac
      841 gcacttggaa tgacccttca gtgcagcagg acatcaagtt cttgcctttc aaggtggttg
      901 aaaagaaaac taaaccatac attcaagttg atattggagg tgggcaaacc aaaacatttg
      961 ccccagaaga aatttctgcc atggttctca ctaaaatgaa agaaactgct gaagcatatt
     1021 tgggaaagaa ggttacccat gcagttgtta ctgtgccggc ttacttcaat gatgcccagc
     1081 gccaagcaac caaagatgct ggcaccattg ctggactgaa tgtcatgcgg atcatcaatg
     1141 agcccacagc agctgctatt gcgtatggcc tggataagag agagggcgag aagaacatcc
     1201 tcgtttttga cctgggcggt ggaaccttcg atgtgtctct tctgaccatt gacaatggtg
     1261 tctttgaagt ggtggccacg aatggagaca ctcatctcgg tggggaagac tttgatcagc
     1321 gggttatgga acacttcatc aagctgtaca aaaagaaaac tgggaaagac gttagaaaag
     1381 acaacagagc tgtgcagaaa cttcgtcgtg aggtggaaaa ggctaagcga gccctgtctt
     1441 ctcagcatca agcaagaatt gagatagagt ccttctttga aggagaagac ttctctgaga
     1501 ccctgactcg ggccaaattt gaagagttga acatggacct gttccgatct accatgaagc
     1561 cagtccagaa agtgttggaa gactctgatc tgaagaaatc agacattgat gaaattgttc
     1621 ttgtcggtgg gtctactcgg attcccaaga ttcagcagct ggtgaaagag ttcttcaatg
     1681 gcaaggagcc atcccgtggc ataaacccag atgaggctgt agcatacggt gctgctgtcc
     1741 aggctggtgt cctctctggt gatcaagata caggtgatct ggtactgctt gatgtatgtc
     1801 ctcttacact tggtattgaa acagtgggag gtgtcatgac caaactgatt ccaaggaaca
     1861 ctgtggtacc caccaagaag tctcagatct tttccacagc ttctgataat cagccaactg
     1921 taacaatcaa ggtctatgaa ggtgaacgac ccctaacaaa agacaaccat cttctgggta
     1981 catttgatct gactggaatt cctcctgctc ctcgtggggt accccagatt gaagtcacct
     2041 ttgagataga tgttaatggt attcttcgag tgacagctga agacaaaggt acagggaaca
     2101 aaaacaaaat cacaattacc aatgaccaaa atcgcctgac acctgaagaa attgaaagga
     2161 tggttaatga tgcagagaag tttgctgagg aagacaaaaa gctcaaagag cgcattgata
     2221 ccaggaacga gttggaaagc tatgcttact ctctcaagaa ccagattgga gataaagaaa
     2281 agctgggcgg taaactttcc tctgaagata aagaaaccat ggagaaagct gtagaggaaa
     2341 agattgaatg gctggaaagc caccaggatg cagacattga agactttaaa gctaaaaaga
     2401 aggaactaga ggaaattgtt cagcctatta ttagcaaact ctatggaagt gcaggccctc
     2461 ccccaactgg tgaagaggat acatcagaag gggatccacc ggtcgccacc atggtgagca
     2521 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     2581 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     2641 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     2701 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     2761 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     2821 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     2881 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     2941 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     3001 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     3061 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     3121 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     3181 gccgccactc caccggcggc atggacgagc tgtacaagaa agacgagcta taagcggccg
     3241 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3301 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3481 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3541 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3601 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3661 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3721 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3781 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3841 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3901 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4081 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4261 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4321 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4381 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4441 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4501 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4561 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4621 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4681 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4741 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4801 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4861 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4921 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4981 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5041 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5101 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5161 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5221 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5281 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5341 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5401 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5461 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5521 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5581 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5641 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5701 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6541 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6601 agttcatagc ccatatatgg agttccg
//
