LOCUS       sequence-458625-        7691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag-tagged wild-type MCRS1 over-expression construct for transient 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  AUTHORS   Mark KG, Kolla S, Aguirre JD, Garshott DM, Schmitt S, Haakonsen DL, 
            Xu C, Kater L, Kempf G, Martinez-Gonzalez B, Akopian D, See SK, 
            Thoma NH, Rape M
  TITLE     Orphan quality control shapes network dynamics and gene expression.
  JOURNAL   Cell. 2023 Aug 3;186(16):3460-3475.e23. doi: 
            10.1016/j.cell.2023.06.015. Epub 2023 Jul 20.
  PUBMED    37478862
REFERENCE   2  (bases 1 to 7691)
  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"; doi: 
            "10.1016/j.cell.2023.06.015"; journalName: "Cell"; date: "2023-08-3-
            3"; volume: "186"; issue: "16"; pages: "3460-3475.e23"
COMMENT     Sequence Label: pCMV 3xFLAG-MCRS1
FEATURES             Location/Qualifiers
     source          1..7691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             956..1021
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2475..3097
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2674..2693)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3121..3141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3139..3436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3490..4284
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3544..3563)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4154..4173
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4936..5070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4973..4992)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5027..5046
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5075..5094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5076..5094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5108..5124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5108..5124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5121..5143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5132..5148
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5156..5162,5163..5180,5181..5186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5201..5222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5339..5356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5510..6098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5590..5609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6269..7060,7061..7129))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6892..6911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7130..7234)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7261..7689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7348..7367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7558..7579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaattaa ccatggacta
      961 caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa
     1021 gcttgcggcc gcgaattcag gccggcccat ggacaaagat tctcaggggc tgctagattc
     1081 atccctgatg gcatcaggca ctgccagccg ctcagaggat gaggagtcac tggcagggca
     1141 gaagcgagcc tcctcccagg ccttgggcac catccctaaa cggagaagct cctccaggtt
     1201 catcaagagg aagaagttcg atgatgagct ggtggagagc agcctggcaa aatcttctac
     1261 ccgggcaaag ggggccagtg gggtggaacc agggcgctgt tcggggagtg aaccctcctc
     1321 cagtgagaag aagaaggtat ccaaagcccc cagcactcct gtgccaccca gcccagcccc
     1381 agcccctgga ctcaccaagc gtgtgaagaa gagtaaacag ccacttcagg tgaccaagga
     1441 tctgggccgc tggaagcctg cagatgacct cctgctcata aatgctgtgt tgcagaccaa
     1501 cgacctgacc tccgtccacc tgggcgtgaa attcagctgc cgcttcaccc ttcgggaggt
     1561 ccaggagcgt tggtacgccc tgctctacga tcctgtcatc tccaagttgg cctgtcaggc
     1621 catgaggcag ctgcacccag aggctattgc agccatccag agcaaggccc tgtttagcaa
     1681 ggctgaggag cagctgctga gcaaagtggg atcgaccagc cagcccacct tggagacctt
     1741 ccaggacctg ctgcacagac accctgatgc cttctacctg gcccgtaccg cgaaggccct
     1801 gcaggcccac tggcagctca tgaagcagta ttacctgctg gaggaccaga cagtgcagcc
     1861 gctgcccaaa ggggaccaag tgctgaactt ctctgatgca gaggacctga ttgatgacag
     1921 taagctcaag gacatgcgag atgaggtcct ggaacatgag ctgatggtgg ctgaccggcg
     1981 ccagaagcga gagattcggc agctggaaca ggaactgcat aagtggcagg tgctagtgga
     2041 cagcatcaca ggcatgagct ctccggactt cgacaaccag acactggcag tgctgcgggg
     2101 ccgcatggtg cggtacctga tgcgctcgcg tgagatcacc ctgggcagag caaccaagga
     2161 taaccagatt gatgtggacc tgtctctgga gggtccggcc tggaagatat cccggaaaca
     2221 aggtgtcatc aagctgaaga acaacggtga tttcttcatt gccaatgagg gtcgacggcc
     2281 catctacatc gatggacggc cggtgctctg tggctccaaa tggcgcctca gcaacaactc
     2341 tgtggtggag atcgccagcc tgcgattcgt cttccttatc aaccaggacc tcattgccct
     2401 catcagggct gaggctgcca agatcacacc acagtgaggc gcgccttcgg taccagtcga
     2461 ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     2521 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     2581 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     2641 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     2701 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     2761 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     2821 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     2881 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     2941 tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag acacagcata
     3001 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct
     3061 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt
     3121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3361 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3421 gaggaggctt ttttggagga attgatcagc ttgggatctg atcaagagac aggatgagga
     3481 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3541 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3601 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3661 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg cgacgacggg cgttccttgc
     3721 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3781 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3841 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3901 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3961 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4021 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4081 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4141 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4201 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4261 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4321 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4381 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4441 agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc
     4501 tggacgacct cgcggagttc taccggcagt gcaaatccgt cggcatccag gaaaccagca
     4561 gcggctatcc gcgcatccat gcccccgaac tgcaggagtg gggaggcacg atggccgctt
     4621 tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa cgaattgctt gcaggcatct
     4681 catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg tggatggagc gctggcgcct
     4741 gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc tggaaccgta aaaaggccgc
     4801 gttgctggcg tttttccata ggctccgccg atcataatca gccataccac atttgtagag
     4861 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4921 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4981 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5041 ctcatcaatg tatcttatca tgtctggatc aattccctat agtgagtcgt attaaattcg
     5101 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7261 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7321 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7381 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     7441 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     7501 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7561 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7621 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7681 acgcgaattt t
//
