LOCUS       sequence-363551-        4881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red fluorescent reporter for polymerized actin. For in vitro 
            transcription (SP6) or expression through CMV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4881)
  AUTHORS   Boutillon A, Escot S, Elouin A, Jahn D, Gonzalez-Tirado S, Starruss 
            J, Brusch L, David NB
  TITLE     Guidance by followers ensures long-range coordination of cell 
            migration through alpha-catenin mechanoperception.
  JOURNAL   Dev Cell. 2022 May 17. pii: S1534-5807(22)00329-X. doi: 
            10.1016/j.devcel.2022.05.001.
  PUBMED    35613615
REFERENCE   2  (bases 1 to 4881)
  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: "Dev Cell. 
            2022 May 17. pii: S1534-5807(22)00329-X. doi: 
            10.1016/j.devcel.2022.05.001."
COMMENT     Sequence Label: pCS2+ Lifeact-mCherry
FEATURES             Location/Qualifiers
     source          1..4881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..65
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     47..64
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             97..147
                     /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"
     regulatory      178..187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             184..894
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(329..347)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     597..616
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    945..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(969..988)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1023..1042
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1192..1212)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1192..1210)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1231..1247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1231..1247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1244..1266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1255..1271
                     /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(1279..1285,1286..1303,1304..1309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1324..1345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1462..1479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1633..2221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1713..1732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2392..3183,3184..3252))
                     /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     3015..3034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3253..3357)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3383..3838)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3520..3541)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3732..3751
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(3909..4881,1..12)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgtttaggga accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact
       61 atagaataca agctacttgt tctttttgca ggatccatgg gagttgctga tcttattaaa
      121 aaattcgaat caatttcaaa agaagaagga gatccaccag ttgctactca ggatcccgcc
      181 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
      241 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      301 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
      361 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
      421 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      481 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      541 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
      601 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
      661 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
      721 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
      781 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
      841 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc
      901 aaggcctctc gagcctctag aactatagtg agtcgtatta cgtagatcca gacatgataa
      961 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1021 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1081 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1141 aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     1201 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1261 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     1321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3361 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     3421 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3481 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3541 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3601 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3661 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3721 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3781 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     3841 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     3901 ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     3961 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     4021 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
     4081 cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
     4141 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
     4201 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
     4261 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
     4321 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
     4381 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
     4441 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
     4501 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
     4561 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
     4621 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
     4681 tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt
     4741 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
     4801 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
     4861 gaggtctata taagcaatgc t
//
