LOCUS       sequence-227428-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfCherry11x4-ß-actin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  AUTHORS   Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert LA, 
            Ishikawa H, Leonetti MD, Marshall WF, Weissman JS, Huang B
  TITLE     Versatile protein tagging in cells with split fluorescent protein.
  JOURNAL   Nat Commun. 2016 Mar 18;7:11046. doi: 10.1038/ncomms11046.
  PUBMED    26988139
REFERENCE   2  (bases 1 to 5412)
  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: "Nat 
            Commun."; date: "2016-03-18"; volume: "7:11046. doi"; pages: " 
            10.1038/ncomms11046"
COMMENT     Sequence Label: pEGFP-sfCherry11x4-ß-actin
FEATURES             Location/Qualifiers
     source          1..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2207..2328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2244..2263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2298..2317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2335..2790)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2472..2493)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2684..2703
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2817..2921
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2923..3280
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2923..3114
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2946..2966)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3131..3266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3315..4109
                     /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             3315..4109
                     /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(3369..3388)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3979..3998
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4297..4316)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4341..4388
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4717..5305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5206..5225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tgtacacaat cgttgaacag tacgagaggg cagagggtag
      661 gcatagcaca ggtggagggg gcagtggcgg ctacaccatc gtcgagcaat atgagagagc
      721 tgaaggccgc cactctacag gtggaggagg ttctggtggg tataccatcg ttgaacagta
      781 cgaacgcgct gagggcaggc actccacagg aggtggtggt agtggggggt ataccatcgt
      841 cgagcagtac gagcgcgccg agggcaggca cagtaccggt ggtggaggat ctggcgggtc
      901 cggactcaga tctggcagcg gtggaggcag cgcatccggc ggaagcggaa gcgatgatga
      961 tatcgccgcg ctcgtcgtcg acaacggctc cggcatgtgc aaggccggct tcgcgggcga
     1021 cgatgccccc cgggccgtct tcccctccat cgtggggcgc cccaggcacc agggcgtgat
     1081 ggtgggcatg ggtcagaagg attcctatgt gggcgacgag gcccagagca agagaggcat
     1141 cctcaccctg aagtacccca tcgagcacgg catcgtcacc aactgggacg acatggagaa
     1201 aatctggcac cacaccttct acaatgagct gcgtgtggct cccgaggagc accccgtgct
     1261 gctgaccgag gcccccctga accccaaggc caaccgcgag aagatgaccc agatcatgtt
     1321 tgagaccttc aacaccccag ccatgtacgt tgctatccag gctgtgctat ccctgtacgc
     1381 ctctggccgt accactggca tcgtgatgga ctccggtgac ggggtcaccc acactgtgcc
     1441 catctacgag gggtatgccc tcccccatgc catcctgcgt ctggacctgg ctggccggga
     1501 cctgactgac tacctcatga agatcctcac cgagcgcggc tacagcttca ccaccacggc
     1561 cgagcgggaa atcgtgcgtg acattaagga gaagctgtgc tacgtcgccc tggacttcga
     1621 gcaagagatg gccacggctg cttccagctc ctccctggag aagagctacg agctgcctga
     1681 cggccaggtc atcaccattg gcaatgagcg gttccgctgc cctgaggcac tcttccagcc
     1741 ttccttcctg ggcatggagt cctgtggcat ccacgaaact accttcaact ccatcatgaa
     1801 gtgtgacgtg gacatccgca aagacctgta cgccaacaca gtgctgtctg gcggcaccac
     1861 catgtaccct ggcattgccg acaggatgca gaaggagatc actgccctgg cacccagcac
     1921 aatgaagatc aagatcattg ctcctcctga gcgcaagtac tccgtgtgga tcggcggctc
     1981 catcctggcc tcgctgtcca ccttccagca gatgtggatc agcaagcagg agtatgacga
     2041 gtccggcccc tccatcgtcc accgcaaatg cttctaggga tccaccggat ctagataact
     2101 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2161 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2221 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2281 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     2341 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2401 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2461 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2521 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2581 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2641 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2701 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2761 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     2821 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2881 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     2941 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3001 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3121 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3181 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3241 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     3301 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3361 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3421 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3481 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3541 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3601 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3661 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3721 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3781 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3841 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3901 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3961 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4021 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4081 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4141 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4201 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4261 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     4321 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     4381 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     4441 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     4501 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     4561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5341 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5401 ccgtattacc gc
//
