LOCUS       sequence-258411-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes Thy1.1 followed by a P2A site and muSOX D219A catalytic 
            mutant for enriching pure populations of transfected cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  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 5949)
  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: Thy1.1-muSOX D219A
FEATURES             Location/Qualifiers
     source          1..5949
                     /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             598..1083
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     polyA_signal    2737..2858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2828..2847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2865..3320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3002..3023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3214..3233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3347..3451
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3453..3810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3453..3644
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3476..3496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3661..3796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3845..4639
                     /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             3845..4639
                     /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(3899..3918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4509..4528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4827..4846)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4871..4918
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5247..5835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5736..5755
                     /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 gatgggtatg
      601 aacccagcca tcagcgtcgc tctcctgctc tcagtcttgc aggtgtcccg agggcagaag
      661 gtgaccagcc tgacagcctg cctggtgaac caaaaccttc gcctggactg ccgccatgag
      721 aataacacca aggataactc catccagcat gagttcagcc tgacccgaga gaagaggaag
      781 cacgtgctct caggcaccct tgggataccc gagcacacgt accgctcccg cgtcaccctc
      841 tccaaccagc cctatatcaa ggtccttacc ctagccaact tcaccaccaa ggatgagggc
      901 gactactttt gtgagcttcg cgtctcgggc gcgaatccca tgagctccaa taaaagtatc
      961 agtgtgtata gagacaagct ggtcaagtgt ggcggcataa gcctgctggt tcagaacaca
     1021 tcctggatgc tgctgctgct gctttccctc tccctcctcc aagccctgga cttcatttct
     1081 ctgaacttcg accttcttaa gcttgccgga gacgtcgagt ccaaccctgg gcccttggga
     1141 attctcatgg aagggtcgat tattctggat ttttttgaaa aggatgattt gttggaagtt
     1201 ctgggaaaca tggcccggga aaatcaacta gaatcactgg caactcttaa tttttctgca
     1261 tttataagat caccccaagt tcaagagctc ctggcaacat caaagacttc tgtacgtatt
     1321 tctgatatga ggttgacgta ctattacttt ctgtttctca gacttaatga atatattggg
     1381 aatactgcca taatgggcgt ttttaaggac atgatgcacc tgacagacaa cagtgaggtg
     1441 tctgccgtgt atgctgcgtg tcgggacgtg accccagata taaagtatgc agtgtgccag
     1501 agaatagaag ccctgactag gggacagcag gataatgaac tatgggacat tctgcgtgac
     1561 ggcatgatct catcatctaa atttcagtgg gctgtaaaac aacatagcat caacaagaaa
     1621 ctgtttaatc cacagcctat aaaggttaat cactattttg ctggtcctct agcctttggg
     1681 ataagatgtg aacaaactgt taaaaaaatt ttgagtgagc ttattcatcc aaacttgcca
     1741 agatttcatg attgtggatt tttgcccagt gcaattgatg gaatttttgg agtttccctg
     1801 gccactgctt tcaatgtgtt tacagatagc agcggacttg tgcactttga accagatagt
     1861 attgtgtatg aaataaagtc cagatataag taccagtttt caaaatctga atttgatggt
     1921 ctggccaaga aatatttgga tctctacaaa aacccctgtg aggcaacttt tatcaagttc
     1981 ataaactgta tttcaagacc cgctgttgaa tatgttccac atggaaaact tccctcagaa
     2041 agtgactatc taatcaccca ctccatgaac tggaaaacca gcaacaagag aaagcgcaaa
     2101 atcactgata gtcatatctg tcttaaaaaa tgtctacttc acaacatgta tcaacaatca
     2161 acagtttata tcttgtcaga cccctctgag acttctggaa aaatcaccat taaggcatct
     2221 tttcccatag atgtatttgt gaatcctgca cacaactact tttaccaggt ggcgctccaa
     2281 aatatggttg ttcaagatta cattgaattt ggtcagggtg tatataaaag actaggccat
     2341 cagaagaatt tcatagcatc tggatttttc agaaagagac acttttctga tcctgctgtt
     2401 tgctccattg gtaattgtgg aaagttggat actacagacg agataccagt ggcgctcatc
     2461 atcactcctg ttaggattcc atctactgtg ctgcacgagt acctcaagaa agcaattgat
     2521 ttttggaatc agtgtgctga ggcaaacttt gaccatatcc catgggggca gctttcggat
     2581 gttgcaagaa aacccataac cccctaagga tccaccggat ctagataact gatcataatc
     2641 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2701 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2821 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2881 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2941 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3001 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3061 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3121 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3181 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3241 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3301 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3421 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3721 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3781 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3841 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3901 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3961 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4021 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4081 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4141 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4201 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4261 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4321 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4381 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4441 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4501 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4561 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4621 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4681 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4801 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4861 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4921 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4981 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5041 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5101 atactttaga ttgatttaaa acttcatttt taatttaaaa aaatctaggt gaagatcctt
     5161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5221 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5281 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5341 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5401 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5461 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5521 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5581 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5641 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5701 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5761 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5821 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5881 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5941 gccatgcat
//
