LOCUS       sequence-264393-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Drosophila melanogaster Cofilin (Twinstar) in bacterial 
            cell culture.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  AUTHORS   Figard L, Zheng L, Biel N, Xue Z, Seede H, Coleman S, Golding I, 
            Sokac AM
  TITLE     Cofilin-Mediated Actin Stress Response Is Maladaptive in 
            Heat-Stressed Embryos.
  JOURNAL   Cell Rep. 2019 Mar 26;26(13):3493-3501.e4. doi: 
            10.1016/j.celrep.2019.02.092.
  PUBMED    30917306
REFERENCE   2  (bases 1 to 5647)
  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: 
            "10.1016/j.celrep.2019.02"; journalName: "Cell Rep"; date: 
            "2019-03-26- 26"; volume: "26"; issue: "13"; pages: "3493-3501.e4"
COMMENT     Sequence Label: pET45b-tsr(B/X) (6X-His-Dm-Cofilin)
FEATURES             Location/Qualifiers
     source          1..5647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             43..57
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             526..570
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(608..626)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      622..669
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      706..1161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(793..812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1003..1024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1276..2067,2068..2136))
                     /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     1899..1918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2137..2229)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2310..2898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2799..2818
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3052..3069
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3084..3223
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3149..3168)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3309..3331
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3325..3516)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4088..5170)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5132..5151
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5171..5248)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5454..5473
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5561..5580
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5561..5579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5580..5604
                     /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)."
     RBS             5619..5641
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga cgacaagagt
       61 ccggatcccg cttctggtgt aactgtgtct gatgtctgca agactacata cgaggaaata
      121 aagaaggaca aaaagcatcg ctatgtcatc ttttacattc gcgatgaaaa gcagattgat
      181 gtggagactg tggcagatcg caacgccgag tacgaccagt ttctagaaga tatccagaaa
      241 tgcggacctg gagagtgccg gtacggactg ttcgatttcg agtacatgca ccagtgccaa
      301 ggcacctctg agagttcaaa gaagcaaaag ctgttcctta tgtcgtggtg tcccgatact
      361 gccaaggtca agaagaagat gttgtactcc agctccttcg atgctctcaa gaagtcgctc
      421 gtgggcgtcc aaaagtatat acaagccact gatctttccg aagcctcccg ggaagccgtc
      481 gaggagaaac tccgggccac cgaccgccaa taactcgagt ctggtaaaga aaccgctgct
      541 gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct
      601 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      661 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
      721 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      781 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      841 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      901 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      961 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1021 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1081 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1141 aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa aaaggatctt
     1201 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1261 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1321 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1381 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1441 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1501 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1561 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1621 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1681 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1741 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1801 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1861 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1921 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1981 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2041 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2101 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatcatgatt
     2161 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2221 ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2281 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2341 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2401 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2461 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2521 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2581 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2641 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2701 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2761 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2821 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2881 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2941 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3001 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3061 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3121 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3181 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3241 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3301 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3361 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3421 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3481 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     3541 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3601 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3661 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3721 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     3781 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     3841 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     3901 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     3961 gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4021 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4201 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4261 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4321 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     4381 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     4441 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4501 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     4561 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     4621 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     4681 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     4741 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     4801 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     4861 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     4921 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     4981 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5041 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5101 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     5161 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5221 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     5281 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     5341 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     5401 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     5461 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     5521 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg
     5581 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag
     5641 atatacc
//
