LOCUS       sequence-167858-        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-S15D/S82D
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1658..2280
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1857..1876)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2304..2324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2309..2638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2489..2624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2676..2695)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2677..2695)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2709..2725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2709..2725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2722..2744)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2733..2749
                     /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(2757..2763,2764..2781,2782..2787))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2802..2823
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2940..2957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3111..3699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3191..3210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3870..4661,4662..4730))
                     /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     4493..4512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4731..4835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4862..5290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4949..4968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5159..5180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgcggccgcg aattcatcga tagatctgat atcggtacca gtcgactcta gaggatccag
     1021 catgaccgag cgccgcgtcc ccttctcgct cctgcggggc cccgattggg accccttccg
     1081 cgactggtac ccgcatagcc gcctcttcga ccaggccttc gggctgcccc ggctgccgga
     1141 ggagtggtcg cagtggttag gcggcagcag ctggccaggc tacgtgcgcc ccctgccccc
     1201 cgccgccatc gagagccccg cagtggccgc gcccgcctac agccgcgcgc tcagccggca
     1261 actcgatagc ggggtctcgg agatccggca cactgcggac cgctggcgcg tgtccctgga
     1321 tgtcaaccac ttcgccccgg acgagctgac ggtcaagacc aaggatggcg tggtggagat
     1381 caccggcaag cacgaggagc ggcaggacga gcatggctac atctcccggt gcttcacgcg
     1441 gaaatacacg ctgccccccg gtgtggaccc cacccaagtt tcctcctccc tgtcccctga
     1501 gggcacactg accgtggagg cccccatgcc caagctagcc acgcagtcca acgagatcac
     1561 catcccagtc accttcgagt cgcgggccca gcttgggggc ccagaagctg caaaatccga
     1621 tgagactgcc gccaagtaaa gccttagccc ggatcccggg tggcatccct gtgacccctc
     1681 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     1741 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     1801 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     1861 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     1921 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     1981 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     2041 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     2101 acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca
     2161 ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag
     2221 ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt
     2281 gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2461 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2521 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2581 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2641 gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc
     2701 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2761 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2821 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2881 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2941 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3001 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3061 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3121 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3181 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3241 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3301 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3361 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3421 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3481 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3541 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3601 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3661 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3721 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3781 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3841 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3901 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3961 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4021 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4081 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4141 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4201 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4261 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4321 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4381 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4441 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4501 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4561 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4621 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4681 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4741 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4801 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5281 aacgcgaatt ttaacaaaat attaacgctt acaattt
//
