LOCUS       sequence-258439-        6220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses muSOX with GFP N-terminally fused.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6220)
  AUTHORS   Gilbertson S, Federspiel JD, Hartenian E, Cristea IM, Glaunsinger B
  TITLE     Changes in mRNA abundance drive shuttling of RNA binding proteins, 
            linking cytoplasmic RNA degradation to transcription.
  JOURNAL   Elife. 2018 Oct 3;7. pii: 37663. doi: 10.7554/eLife.37663.
  PUBMED    30281021
REFERENCE   2  (bases 1 to 6220)
  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: "Elife."; 
            date: "2018-10-3"; volume: "7. pii: 37663. doi"; pages: " 
            10.7554/eLife.37663"
COMMENT     Sequence Label: GFP-muSOX
FEATURES             Location/Qualifiers
     source          1..6220
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1393..1417
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2849..2873)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3008..3129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3045..3064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3099..3118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3136..3591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3273..3294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3485..3504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3618..3722
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3724..4081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3724..3915
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3747..3767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3932..4067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4116..4910
                     /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             4116..4910
                     /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(4170..4189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4780..4799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5098..5117)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5142..5189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5518..6106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6007..6026
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcgggcccat caacaagttt gtacaaaaaa gttggcatgg aagggtcgat tattctggat
     1441 ttttttgaaa aggatgattt gttggaagtt ctgggaaaca tggcccggga aaatcaacta
     1501 gaatcactgg caactcttaa tttttctgca tttataagat caccccaagt tcaagagctc
     1561 ctggcaacat caaagacttc tgtacgtatt tctgatatga ggttgacgta ctattacttt
     1621 ctgtttctca gacttaatga atatattggg aatactgcca taatgggcgt ttttaaggac
     1681 atgatgcacc tgacagacaa cagtgaggtg tctgccgtgt atgctgcgtg tcgggacgtg
     1741 accccagata taaagtatgc agtgtgccag agaatagaag ccctgactag gggacagcag
     1801 gataatgaac tatgggacat tctgcgtgac ggcatgatct catcatctaa atttcagtgg
     1861 gctgtaaaac aacatagcat caacaagaaa ctgtttaatc cacagcctat aaaggttaat
     1921 cactattttg ctggtcctct agcctttggg ataagatgtg aacaaactgt taaaaaaatt
     1981 ttgagtgagc ttattcatcc aaacttgcca agatttcatg attgtggatt tttgcccagt
     2041 gcaattgatg gaatttttgg agtttccctg gacactgctt tcaatgtgtt tacagatagc
     2101 agcggacttg tgcactttga accagatagt attgtgtatg aaataaagtc cagatataag
     2161 taccagtttt caaaatctga atttgatggt ctggccaaga aatatttgga tctctacaaa
     2221 aacccctgtg aggcaacttt tatcaagttc ataaactgta tttcaagacc cgctgttgaa
     2281 tatgttccac atggaaaact tccctcagaa agtgactatc taatcaccca ctccatgaac
     2341 tggaaaacca gcaacaagag aaagcgcaaa atcactgata gtcatatctg tcttaaaaaa
     2401 tgtctacttc acaacatgta tcaacaatca acagtttata tcttgtcaga cccctctgag
     2461 acttctggaa aaatcaccat taaggcatct tttcccatag atgtatttgt gaatcctgca
     2521 cacaactact tttaccaggt ggcgctccaa aatatggttg ttcaagatta cattgaattt
     2581 ggtcagggtg tatataaaag actaggccat cagaagaatt tcatagcatc tggatttttc
     2641 agaaagagac acttttctga tcctgctgtt tgctccattg gtaattgtgg aaagttggat
     2701 actacagacg agataccagt ggcgctcatc atcactcctg ttaggattcc atctactgtg
     2761 ctgcacgagt acctcaagaa agcaattgat ttttggaatc agtgtgctga ggcaaacacg
     2821 cttaatttgc tttaattact ttaataatac ccaactttct tgtacaaagt ggttgatggg
     2881 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2941 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3001 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3121 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3181 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3241 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3301 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3361 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3421 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3481 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3541 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3601 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3661 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3721 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3901 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3961 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4021 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4081 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4141 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4201 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4261 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4321 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4381 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4441 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4501 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4561 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4621 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4681 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4741 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4801 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4861 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4921 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4981 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5041 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5101 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5161 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5221 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5281 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5341 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5401 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5461 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5521 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5581 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5641 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5701 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5761 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5821 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5881 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5941 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6001 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6061 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6181 atcccctgat tctgtggata accgtattac cgccatgcat
//
