LOCUS       sequence-314151-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes hamster BiP/GRP78/HSPA5 fused to monomeric EGFP. 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 6633)
  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 6633)
  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-mGFP
FEATURES             Location/Qualifiers
     source          1..6633
                     /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             join(2511..2513,2514..2516,2517..3227)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2556..2577)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2817..2836)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3164..3185
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3361..3482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3398..3417)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3452..3471
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3489..3944)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3626..3647)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3838..3857
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3971..4075
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4077..4434
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4077..4268
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4100..4120)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4285..4420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4469..5263
                     /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             4469..5263
                     /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(4523..4542)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5133..5152
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5451..5470)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5495..5542
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5871..6459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6360..6379
                     /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 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2581 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2641 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2701 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2761 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2821 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2881 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2941 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     3001 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3061 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3121 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3181 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggat gaattgtaag
     3241 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3301 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3361 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3421 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3481 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3541 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3601 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3661 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3721 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3781 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3841 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3901 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3961 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4021 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     4081 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4141 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4201 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4261 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4321 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4381 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4441 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4501 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4561 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4621 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4681 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4741 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4801 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4861 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4921 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4981 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5041 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5101 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5161 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5221 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5281 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5341 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5401 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5461 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5521 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5581 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5641 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5701 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5761 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5821 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6481 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6541 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     6601 gtcattagtt catagcccat atatggagtt ccg
//
