LOCUS       sequence-173286-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hsp27 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  TITLE     Moens lab Hsp27 plasmids
REFERENCE   2  (bases 1 to 5317)
  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"
COMMENT     Sequence Label: pFLAG-CMV2-Hsp27-S78D/S82D
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..67
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     59..76
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     60..76
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             854..877
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1577..2199
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1776..1795)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2223..2243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2228..2557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2408..2543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2595..2614)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2596..2614)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2628..2644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2628..2644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2641..2663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2652..2668
                     /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(2676..2682,2683..2700,2701..2706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2721..2742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2859..2876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3030..3618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3110..3129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3789..4580,4581..4649))
                     /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     4412..4431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4650..4754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4781..5236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4868..4887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5078..5099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
       61 taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc aattagccat
      121 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
      181 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggta
      721 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagaa
      841 ttgatctacc atggactaca aagacgatga cgacaagctt gcggccgcga attcatcgat
      901 agatctgata tcggtaccag tcgactctag aggatccagc atgaccgagc gccgcgtccc
      961 cttctcgctc ctgcggggcc ccagctggga ccccttccgc gactggtacc cgcatagccg
     1021 cctcttcgac caggccttcg ggctgccccg gctgccggag gagtggtcgc agtggttagg
     1081 cggcagcagc tggccaggct acgtgcgccc cctgcccccc gccgccatcg agagccccgc
     1141 agtggccgcg cccgcctaca gccgcgcgct cgatcggcaa ctcgatagcg gggtctcgga
     1201 gatccggcac actgcggacc gctggcgcgt gtccctggat gtcaaccact tcgccccgga
     1261 cgagctgacg gtcaagacca aggatggcgt ggtggagatc accggcaagc acgaggagcg
     1321 gcaggacgag catggctaca tctcccggtg cttcacgcgg aaatacacgc tgccccccgg
     1381 tgtggacccc acccaagttt cctcctccct gtcccctgag ggcacactga ccgtggaggc
     1441 ccccatgccc aagctagcca cgcagtccaa cgagatcacc atcccagtca ccttcgagtc
     1501 gcgggcccag cttgggggcc cagaagctgc aaaatccgat gagactgccg ccaagtaaag
     1561 ccttagcccg gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     1621 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     1681 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     1741 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     1801 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     1861 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     1921 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     1981 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     2041 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     2101 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     2161 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     2221 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2281 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2401 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2461 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2521 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa
     2581 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
     2641 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2701 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2761 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2821 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2881 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2941 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4741 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     4801 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4861 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4921 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4981 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5041 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5101 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5161 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5221 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5281 tgcgggcctc ttcgctatta cgccagctgg cgaaagg
//
