LOCUS       sequence-461409-        6587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human beta actin fused intramolecularly to 
            monomeric superfolder GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6587)
  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 6587)
  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-beta-actin
FEATURES             Location/Qualifiers
     source          1..6587
                     /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(2729..2746)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2735..2959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3092..3111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3243..3290
                     /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    4029..4150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4066..4085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4120..4139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4199..4215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4199..4215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4212..4234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4223..4239
                     /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(4247..4253,4254..4271,4272..4277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4292..4313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4430..4447)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4601..5189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4681..4700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5360..6151,6152..6220))
                     /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     5983..6002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6221..6325)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6400..6419)
                     /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 ggatgatgat atcgccgcgc
      781 tcgtcgtcga caacggctcc ggcatgtgca aggccggctt cgcgggcgac gatgcccccc
      841 gggccgtctt cccctccatc gtggggcgcc ccaggcacca gggcgtgatg gtgggcatgg
      901 gtcagaagga ttcctatgtg ggcgacgagg cccagagcaa gagaggcatc ctcaccctga
      961 agtaccccat cgagcacggc atcgtcacca actgggacga catggagaaa atctggcacc
     1021 acaccttcta caatgagctg cgtgtggctc ccgaggagca ccccgtgctg ctgaccgagg
     1081 cccccctgaa ccccaaggcc aaccgcgaga agatgaccca gatcatgttt gagaccttca
     1141 acaccccagc catgtacgtt gctatccagg ctgtgctatc cctgtacgcc tctggccgta
     1201 ccactggcat cgtgatggac tccggtgacg gggtcaccca cactgtgccc atctacgagg
     1261 ggtatgccct cccccatgcc atcctgcgtc tggacctggc tggccgggac ctgactgact
     1321 acctcatgaa gatcctcacc gagcgcggct acagcttcac caccacggcc gagcgggaaa
     1381 tcgtgcgtga cattaaggag aagctgtgct acgtcgccct ggacttcgag caagagatgg
     1441 ccacggctgc ttccagcagc ggcagcagcg tgtccaaggg cgaggagctg ttcaccggcg
     1501 tggtgcccat cctggtggag ctggatggcg acgtgaacgg ccacaagttc agcgtgcgcg
     1561 gcgagggcga gggcgacgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     1621 gcaagctgcc cgtgccctgg cccaccctgg tgaccaccct gacctacggc gtgcagtgct
     1681 tcagccgcta ccccgatcac atgaagcagc acgatttctt caagagcgcc atgcccgagg
     1741 gctacgtgca ggagcgcacc atcagcttca aggatgacgg cacctacaag acccgcgccg
     1801 aggtgaagtt cgagggcgat accctggtga accgcatcga gctgaagggc atcgatttca
     1861 aggaggatgg caacatcctg ggccacaagc tggagtacaa cttcaacagc cacaacgtgt
     1921 acatcaccgc cgataagcag aagaacggca tcaaggccaa cttcaagatc cgccacaatg
     1981 tggaggatgg ctccgtgcag ctggccgatc actaccagca gaacaccccc atcggcgacg
     2041 gcccagtgct gctgcccgat aaccactacc tgagcaccca gagcaagctg tccaaggacc
     2101 ccaacgagaa gcgcgatcac atggtgctgc tggagttcgt gaccgccgcc ggcatcaccc
     2161 tgggcatgga tgagctgtac aagagcggcg cgccgggctc ctccctggag aagagctacg
     2221 agctgcctga cggccaggtc atcaccattg gcaatgagcg gttccgctgc cctgaggcac
     2281 tcttccagcc ttccttcctg ggcatggagt cctgtggcat ccacgaaact accttcaact
     2341 ccatcatgaa gtgtgacgtg gacatccgca aagacctgta cgccaacaca gtgctgtctg
     2401 gcggcaccac catgtaccct ggcattgccg acaggatgca gaaggagatc actgccctgg
     2461 cacccagcac aatgaagatc aagatcattg ctcctcctga gcgcaagtac tccgtgtgga
     2521 tcggcggctc catcctggcc tcgctgtcca ccttccagca gatgtggatc agcaagcagg
     2581 agtatgacga gtccggcccc tccatcgtcc accgcaaatg cttctgagga tccactagta
     2641 acggccgcca gtgtgctgga attctgcaga tatccatcac actggcggcc gctcgagtct
     2701 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2761 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2821 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2881 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2941 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3001 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3061 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3121 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3181 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc
     3241 tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc cttggccact
     3301 gaatacaaac ccactgtccg actggcaaca agagatgatg tcccaagagc cgtgagaaca
     3361 ctggcagcag ctttcgccga ctacccagcc acaagacaca ctgtggaccc cgatcggcat
     3421 atcgagagag tgaccgagct gcaggaactg ttcctgacaa gggtgggcct ggacattggg
     3481 aaggtctggg tcgcagacga tggagcagct gtggctgtct ggaccacacc agagagcgtg
     3541 gaagccggag ctgtctttgc agagatcggc cctcgaatgg cagaactgag cggctcccgg
     3601 ctggcagcac agcagcagat ggaggggctg ctggccccac accgccctaa ggaaccagca
     3661 tggttcctgg ccactgtggg agtctctccc gatcatcagg gcaaagggct gggaagtgct
     3721 gtggtcctgc ctggggtgga ggctgcagaa cgagctggag tcccagcatt tctggagaca
     3781 tctgccccca ggaacctgcc tttctatgaa cgcctgggct ttactgtgac cgccgatgtg
     3841 gaagtgcccg agggaccccg aacctggtgt atgacccgaa agcctggagc ctaatagagc
     3901 acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3961 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4021 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4081 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4141 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4201 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4261 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4321 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4381 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4441 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4501 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4561 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4621 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4681 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4741 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4801 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4861 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4921 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4981 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5041 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5101 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5161 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5221 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5281 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5341 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5401 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5461 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5521 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5581 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5641 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5701 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5761 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5821 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5881 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5941 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6001 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6061 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6121 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6181 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6241 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6301 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6361 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     6421 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     6481 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     6541 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgc
//
