LOCUS       sequence-433876-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of actin cytoskeleton.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4913)
  TITLE     LifeAct Fluorescent Proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4913)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Lifeact-LSSmOrange
FEATURES             Location/Qualifiers
     source          1..4913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..51
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             70..780
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     483..502
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    840..974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(864..883)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     918..937
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1087..1107)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1087..1105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1126..1142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1126..1142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1139..1161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1150..1166
                     /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(1174..1180,1181..1198,1199..1204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1219..1240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1357..1374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1528..2116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1608..1627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2287..3078,3079..3147))
                     /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     2910..2929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3148..3252)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3278..3733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3415..3436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3627..3646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3804..4788
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        4823..4841
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4823..4840
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atgggtgttg cagacctaat aaagaaattc gagtcaatct cgaaagagga gggttccgga
       61 ccatccggaa tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg
      121 cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
      181 ggcgagggcc gcccctacga gggctttcag accgttaagc tgaaggtgac caagggtggc
      241 cccctgccct tcgcctggga catcttgtcc cctcagttca cctacggctc caaggcctac
      301 gtgaagcacc ccgccgacat ccccgactac ctcaagctgt ccttccccga gggcttcaag
      361 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgactca ggactcctcc
      421 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
      481 ggccccgtaa tgcagaagaa gaccatgggc atggaggcct cctccgagcg gatgtacccc
      541 gaggacggcg ccctgaaggg cgaggacaag ctcaggctga agctgaagga cggcggccac
      601 tacacctccg aggtcaagac cacctacaag gccaagaagc ccgtgcagtt gcccggcgcc
      661 tacatcgtcg acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
      721 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
      781 catcgattgg aattcaaggc ctctcgagcc tctagaacta tagtgagtcg tattacgtag
      841 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      901 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      961 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1021 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     1081 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     1141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     1201 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1321 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     2581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     2641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3241 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     3301 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3361 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3421 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3481 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3541 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3601 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3661 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3721 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     3781 cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat
     3841 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     3901 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     3961 gccaattcaa tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg
     4021 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     4081 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     4141 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     4201 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     4261 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     4321 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     4381 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     4441 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     4501 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     4561 ctattaatag taacttggca agtacattac tattggaagt acgccagggt acattggcag
     4621 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     4681 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     4741 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg ccattctgcc
     4801 tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct acttgttctt
     4861 tttgcaggat ccactagtgg cgcgccatta attaaaggcc ggccagcgat cgc
//
