LOCUS       sequence-161624-        5460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfCherry2(11) tagged keratin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5460)
  AUTHORS   Feng S, Sekine S, Pessino V, Li H, Leonetti MD, Huang B
  TITLE     Improved split fluorescent proteins for endogenous protein labeling.
  JOURNAL   Nat Commun. 2017 Aug 29;8(1):370. doi: 10.1038/s41467-017-00494-8.
  PUBMED    28851864
REFERENCE   2  (bases 1 to 5460)
  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: "2017-08-29"; volume: "8(1):370. doi"; pages: " 
            10.1038/s41467-017-00494-8"
COMMENT     Sequence Label: pEGFP_sfCherry2(11)_Keratin
FEATURES             Location/Qualifiers
     source          1..5460
                     /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"
     polyA_signal    2248..2369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2285..2304)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2339..2358
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2376..2831)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2513..2534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2725..2744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2858..2962
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2964..3321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2964..3155
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2987..3007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3172..3307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3356..4150
                     /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             3356..4150
                     /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(3410..3429)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4020..4039
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4338..4357)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4382..4429
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4758..5346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5247..5266
                     /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 gctagcgcca
      601 ccatgactac ctgcagccgc cagttcacct cctccagctc catgaagggc tcctgcggca
      661 tcgggggcgg catcgggggc ggctccagcc gcatctcctc cgtcctggcc ggagggtcct
      721 gccgcgcccc cagcacctac gggggcggcc tgtctgtctc atcctcccgc ttctcctctg
      781 ggggagccta tgggttgggg ggcggctatg gcggtggctt cagcagcagc agcagcagct
      841 ttggtagtgg ctttggggga ggatatggtg gtggccttgg tgctggcttg ggtggtggct
      901 ttggtggtgg ctttgctggt ggtgatgggc ttctggtggg cagtgagaag gtgaccatgc
      961 agaacctcaa cgaccgcctg gcctcctacc tggacaaggt gcgtgctctg gaggaggcca
     1021 acgccgacct ggaagtgaag atccgtgact ggtaccagag gcagcggcct gctgagatca
     1081 aagactacag tccctacttc aagaccattg aggacctgag gaacaagatt ctcacagcca
     1141 cagtggacaa tgccaatgtc cttctgcaga ttgacaatgc ccgtctggcc gcggatgact
     1201 tccgcaccaa gtatgagaca gagttgaacc tgcgcatgag tgtggaagcc gacatcaatg
     1261 gcctgcgcag ggtgctggac gaactgaccc tggccagagc tgacctggag atgcagattg
     1321 agagcctgaa ggaggagctg gcctacctga agaagaacca cgaggaggag atgaatgccc
     1381 tgagaggcca ggtgggtgga gatgtcaatg tggagatgga cgctgcacct ggcgtggacc
     1441 tgagccgcat tctgaacgag atgcgtgacc agtatgagaa gatggcagag aagaaccgca
     1501 aggatgccga ggaatggttc ttcaccaaga cagaggagct gaaccgcgag gtggccacca
     1561 acagcgagct ggtgcagagc ggcaagagcg agatctcgga gctccggcgc accatgcaga
     1621 acctggagat tgagctgcag tcccagctca gcatgaaagc atccctggag aacagcctgg
     1681 aggagaccaa aggtcgctac tgcatgcagc tggcccagat ccaggagatg attggcagcg
     1741 tggaggagca gctggcccag ctccgctgcg agatggagca gcagaaccag gagtacaaga
     1801 tcctgctgga cgtgaagacg cggctggagc aggagatcgc cacctaccgc cgcctgctgg
     1861 agggcgagga cgcccacctc tcctcctccc agttctcctc tggatcgcag tcatccagag
     1921 atgtgacctc ctccagccgc caaatccgca ccaaggtcat ggatgtgcac gatggcaagg
     1981 tggtgtccac ccacgagcag gtccttcgca ccaagaacag cagcggtgga gcaagcgcag
     2041 ccagtggtag cgcggatcca ccggtcgcca ccatgtacac catcgtggag cagtacgaga
     2101 gagccgaggc cagacacagc acctaagcgg ccgcgactct agatcataat cagccatacc
     2161 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2221 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2281 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2341 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2401 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2461 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2521 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2581 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2641 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2701 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2761 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2821 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2881 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2941 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3001 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3061 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3121 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3301 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3361 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3421 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3481 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3541 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3601 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3661 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3721 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3781 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3841 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3901 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3961 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4021 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4081 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4141 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4201 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4261 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4321 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4381 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4441 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4501 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4561 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4621 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4681 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4741 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4801 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4861 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4921 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4981 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5041 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5101 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5161 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5221 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5281 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5341 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5401 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
