LOCUS       sequence-465705-        6590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human gamma actin fused intramolecularly to 
            monomeric superfolder GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6590)
  AUTHORS   Martins CS, Iv F, Suman SK, Panagiotou TC, Sidor C, Ruso-Lopez M, 
            Plancke CN, Omi S, Pages R, Gomes M, Llewellyn A, Bandi SR, Ramond 
            L, Arbizzani F, Rimoli CV, Schnorrer F, Robin F, Wilde A, LeGoff L, 
            Pedelacq JD, Jegou A, Cabantous S, Rincon SA, Chandre C, Brasselet 
            S, Mavrakis M
  TITLE     Genetically encoded reporters of actin filament organization in 
            living cells and tissues.
  JOURNAL   Cell. 2025 Apr 1:S0092-8674(25)00276-4. doi: 
            10.1016/j.cell.2025.03.003.
  PUBMED    40179884
REFERENCE   2  (bases 1 to 6590)
  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: "Cell. 2025 
            Apr 1:S0092-8674(25)00276-4. doi: 10.1016/j.cell.2025.03.003."
COMMENT     Sequence Label: iGFP-gamma-actin
FEATURES             Location/Qualifiers
     source          1..6590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    589..607
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    610..628
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     638..662
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2732..2749)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2738..2962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3095..3114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3246..3293
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    4032..4153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4069..4088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4123..4142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4202..4218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4202..4218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4215..4237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4226..4242
                     /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(4250..4256,4257..4274,4275..4280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4295..4316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4433..4450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4604..5192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4684..4703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5363..6154,6155..6223))
                     /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     5986..6005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6224..6328)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6403..6422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg
      601 atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga
      661 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      721 gcctccggac tctagcgttt aaacttaagc ttggtaccat gatggaagaa gagatcgccg
      781 cgctggtcat tgacaatggc tccggcatgt gcaaagctgg ttttgctggg gacgacgctc
      841 cccgagccgt gtttccttcc atcgtcgggc gccccagaca ccagggcgtc atggtgggca
      901 tgggccagaa ggactcctac gtgggcgacg aggcccagag caagcgtggc atcctgaccc
      961 tgaagtaccc cattgagcat ggcatcgtca ccaactggga cgacatggag aagatctggc
     1021 accacacctt ctacaacgag ctgcgcgtgg ccccggagga gcacccagtg ctgctgaccg
     1081 aggcccccct gaaccccaag gccaacagag agaagatgac tcagattatg tttgagacct
     1141 tcaacacccc ggccatgtac gtggccatcc aggccgtgct gtccctctac gcctctgggc
     1201 gcaccactgg cattgtcatg gactctggag acggggtcac ccacacggtg cccatctacg
     1261 agggctacgc cctcccccac gccatcctgc gtctggacct ggctggccgg gacctgaccg
     1321 actacctcat gaagatcctc actgagcgag gctacagctt caccaccacg gccgagcggg
     1381 aaatcgtgcg cgacatcaag gagaagctgt gctacgtcgc cctggacttc gagcaggaga
     1441 tggccaccgc cgcatcctcc agcggcagca gcgtgtccaa gggcgaggag ctgttcaccg
     1501 gcgtggtgcc catcctggtg gagctggatg gcgacgtgaa cggccacaag ttcagcgtgc
     1561 gcggcgaggg cgagggcgac gccaccaacg gcaagctgac cctgaagttc atctgcacca
     1621 ccggcaagct gcccgtgccc tggcccaccc tggtgaccac cctgacctac ggcgtgcagt
     1681 gcttcagccg ctaccccgat cacatgaagc agcacgattt cttcaagagc gccatgcccg
     1741 agggctacgt gcaggagcgc accatcagct tcaaggatga cggcacctac aagacccgcg
     1801 ccgaggtgaa gttcgagggc gataccctgg tgaaccgcat cgagctgaag ggcatcgatt
     1861 tcaaggagga tggcaacatc ctgggccaca agctggagta caacttcaac agccacaacg
     1921 tgtacatcac cgccgataag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1981 atgtggagga tggctccgtg cagctggccg atcactacca gcagaacacc cccatcggcg
     2041 acggcccagt gctgctgccc gataaccact acctgagcac ccagagcaag ctgtccaagg
     2101 accccaacga gaagcgcgat cacatggtgc tgctggagtt cgtgaccgcc gccggcatca
     2161 ccctgggcat ggatgagctg tacaagagcg gcgcgccggg ctcttctctg gagaagagct
     2221 acgagctgcc cgatggccag gtcatcacca ttggcaatga gcggttccgg tgtccggagg
     2281 cgctgttcca gccttccttc ctgggtatgg aatcttgcgg catccacgag accaccttca
     2341 actccatcat gaagtgtgac gtggacatcc gcaaagacct gtacgccaac acggtgctgt
     2401 cgggcggcac caccatgtac ccgggcattg ccgacaggat gcagaaggag atcaccgccc
     2461 tggcgcccag caccatgaag atcaagatca tcgcaccccc agagcgcaag tactcggtgt
     2521 ggatcggtgg ctccatcctg gcctcactgt ccaccttcca gcagatgtgg attagcaagc
     2581 aggagtacga cgagtcgggc ccctccatcg tccaccgcaa atgcttctga ggatccacta
     2641 gtaacggccg ccagtgtgct ggaattctgc agatatccat cacactggcg gccgctcgag
     2701 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2761 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2821 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2881 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2941 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3001 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3061 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3121 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3181 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3241 acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc
     3301 actgaataca aacccactgt ccgactggca acaagagatg atgtcccaag agccgtgaga
     3361 acactggcag cagctttcgc cgactaccca gccacaagac acactgtgga ccccgatcgg
     3421 catatcgaga gagtgaccga gctgcaggaa ctgttcctga caagggtggg cctggacatt
     3481 gggaaggtct gggtcgcaga cgatggagca gctgtggctg tctggaccac accagagagc
     3541 gtggaagccg gagctgtctt tgcagagatc ggccctcgaa tggcagaact gagcggctcc
     3601 cggctggcag cacagcagca gatggagggg ctgctggccc cacaccgccc taaggaacca
     3661 gcatggttcc tggccactgt gggagtctct cccgatcatc agggcaaagg gctgggaagt
     3721 gctgtggtcc tgcctggggt ggaggctgca gaacgagctg gagtcccagc atttctggag
     3781 acatctgccc ccaggaacct gcctttctat gaacgcctgg gctttactgt gaccgccgat
     3841 gtggaagtgc ccgagggacc ccgaacctgg tgtatgaccc gaaagcctgg agcctaatag
     3901 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3961 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4021 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4081 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4141 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4201 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4261 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4321 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4381 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6301 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     6361 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     6421 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     6481 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     6541 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
//
