LOCUS       sequence-433879-        4886 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 4886)
  TITLE     LifeAct Fluorescent Proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4886)
  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            "Unpublished"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-mRuby
FEATURES             Location/Qualifiers
     source          1..4886
                     /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..750
                     /codon_start=1
                     /product="monomeric variant of the red fluorescent protein 
                     eqFP611 (Kredel et al., 2009)"
                     /label=mRuby
                     /translation="MNSLIKENMRMKVVLEGSVNGHQFKCTGEGEGNPYMGTQTMRIKV
                     IEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGVITV
                     MQDTSLEDGCLVYHAQVRGVNFPSNGAVMQKKTKGWEPNTEMMYPADGGLRGYTHMALK
                     VDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVAKFA
                     GLGGG"
     polyA_signal    813..947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(837..856)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     891..910
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1060..1080)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1060..1078)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1099..1115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1099..1115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1112..1134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1123..1139
                     /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(1147..1153,1154..1171,1172..1177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1192..1213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1330..1347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1501..2089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1581..1600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2260..3051,3052..3120))
                     /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     2883..2902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3121..3225)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3251..3706)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3388..3409)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3600..3619
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3777..4761
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        4796..4814
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4796..4813
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atgggtgttg cagacctaat aaagaaattc gagtcaatct cgaaagagga gggttccgga
       61 ccatccggaa tgaacagcct gatcaaagaa aacatgcgga tgaaggtggt gctggaaggc
      121 agcgtgaacg gccaccagtt caagtgcacc ggcgagggcg agggcaaccc ctacatgggc
      181 acccagacca tgcggatcaa agtgatcgag ggcggacctc tgcccttcgc cttcgacatc
      241 ctggccacat ccttcatgta cggcagccgg accttcatca agtaccccaa gggcatcccc
      301 gatttcttca agcagagctt ccccgagggc ttcacctggg agagagtgac cagatacgag
      361 gacggcggcg tgatcaccgt gatgcaggac accagcctgg aagatggctg cctggtgtac
      421 catgcccagg tcaggggcgt gaattttccc agcaacggcg ccgtgatgca gaagaaaacc
      481 aagggctggg agcccaacac cgagatgatg taccccgctg acggcggact gagaggctac
      541 acccacatgg ccctgaaggt ggacggcgga gggcacctga gctgcagctt cgtgaccacc
      601 taccgatcca agaaaaccgt gggcaacatc aagatgcccg gcatccacgc cgtggaccac
      661 cggctggaaa ggctggaaga gtccgacaac gagatgttcg tggtgcagcg ggagcacgcc
      721 gtggccaagt tcgccggcct gggcggaggg taacatcgat tggaattcaa ggcctctcga
      781 gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
      841 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      901 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      961 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
     1021 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     1081 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1141 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1201 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1261 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1381 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1441 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1921 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2101 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3241 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3301 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3361 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3421 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3481 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3541 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3601 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3661 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     3721 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
     3781 accgccaatt caatatggcg tatatggact catgccaatt caatatggtg gatctggacc
     3841 tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc
     3901 ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac
     3961 tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc
     4021 ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta
     4081 tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg
     4141 ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct
     4201 attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa
     4261 taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt
     4321 gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg
     4381 gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt
     4441 caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga
     4501 cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat
     4561 tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa
     4621 atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg
     4681 caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata
     4741 agcaatgctc gtttagggaa ccgccattct gcctggggac gtcggagcaa gcttgattta
     4801 ggtgacacta tagaatacaa gctacttgtt ctttttgcag gatccactag tggcgcgcca
     4861 ttaattaaag gccggccagc gatcgc
//
