LOCUS       sequence-363147-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human beta actin tagged with GFP11..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5184)
  AUTHORS   Tenner B, Zhang JZ, Kwon Y, Pessino V, Feng S, Huang B, Mehta S, 
            Zhang J
  TITLE     FluoSTEPs: Fluorescent biosensors for monitoring compartmentalized 
            signaling within endogenous microdomains.
  JOURNAL   Sci Adv. 2021 May 21;7(21). pii: 7/21/eabe4091. doi: 
            10.1126/sciadv.abe4091. Print 2021 May.
  PUBMED    34020947
REFERENCE   2  (bases 1 to 5184)
  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: "Sci Adv."; 
            date: "2021-05-21"; volume: "7(21). pii: 7/21/eabe4091. doi"; pages:
            " 10.1126/sciadv.abe4091. Print 2021 May"
COMMENT     Sequence Label: pEGFP-GFP11-Actin
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             616..663
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     polyA_signal    1972..2093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2009..2028)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2063..2082
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2100..2555)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2237..2258)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2449..2468
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2582..2686
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2688..3045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2688..2879
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2711..2731)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2896..3031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3080..3874
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3080..3874
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3134..3153)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3744..3763
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4062..4081)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4106..4153
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4482..5070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4971..4990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatgcgtga ccacatggtc cttcatgagt atgtaaatgc tgctgggatt
      661 acatccggac tcagatctgg cagcggtgga ggcagcgcat ccggcggaag cggaagcgat
      721 gatgatatcg ccgcgctcgt cgtcgacaac ggctccggca tgtgcaaggc cggcttcgcg
      781 ggcgacgatg ccccccgggc cgtcttcccc tccatcgtgg ggcgccccag gcaccagggc
      841 gtgatggtgg gcatgggtca gaaggattcc tatgtgggcg acgaggccca gagcaagaga
      901 ggcatcctca ccctgaagta ccccatcgag cacggcatcg tcaccaactg ggacgacatg
      961 gagaaaatct ggcaccacac cttctacaat gagctgcgtg tggctcccga ggagcacccc
     1021 gtgctgctga ccgaggcccc cctgaacccc aaggccaacc gcgagaagat gacccagatc
     1081 atgtttgaga ccttcaacac cccagccatg tacgttgcta tccaggctgt gctatccctg
     1141 tacgcctctg gccgtaccac tggcatcgtg atggactccg gtgacggggt cacccacact
     1201 gtgcccatct acgaggggta tgccctcccc catgccatcc tgcgtctgga cctggctggc
     1261 cgggacctga ctgactacct catgaagatc ctcaccgagc gcggctacag cttcaccacc
     1321 acggccgagc gggaaatcgt gcgtgacatt aaggagaagc tgtgctacgt cgccctggac
     1381 ttcgagcaag agatggccac ggctgcttcc agctcctccc tggagaagag ctacgagctg
     1441 cctgacggcc aggtcatcac cattggcaat gagcggttcc gctgccctga ggcactcttc
     1501 cagccttcct tcctgggcat ggagtcctgt ggcatccacg aaactacctt caactccatc
     1561 atgaagtgtg acgtggacat ccgcaaagac ctgtacgcca acacagtgct gtctggcggc
     1621 accaccatgt accctggcat tgccgacagg atgcagaagg agatcactgc cctggcaccc
     1681 agcacaatga agatcaagat cattgctcct cctgagcgca agtactccgt gtggatcggc
     1741 ggctccatcc tggcctcgct gtccaccttc cagcagatgt ggatcagcaa gcaggagtat
     1801 gacgagtccg gcccctccat cgtccaccgc aaatgcttct agggatccac cggatctaga
     1861 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1921 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2041 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     2101 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2161 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2221 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2281 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2341 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2401 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2461 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2521 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2581 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2641 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2701 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2761 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2821 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2881 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2941 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3001 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3061 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3121 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3181 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3241 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3301 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3361 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3421 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3481 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3541 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3601 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3661 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3721 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3781 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3841 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3901 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3961 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4021 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4081 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4141 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4201 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4261 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4321 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4381 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4621 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5101 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5161 gataaccgta ttaccgccat gcat
//
