LOCUS       sequence-409602-        7279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express fluorescently labeled MS2 coat binding protein (MCP) 
            fused with RNA binding protein TIAL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7279)
  AUTHORS   Horste EL, Fansler MM, Cai T, Chen X, Mitschka S, Zhen G, Lee FCY, 
            Ule J, Mayr C
  TITLE     Subcytoplasmic location of translation controls protein output.
  JOURNAL   Mol Cell. 2023 Dec 21;83(24):4509-4523.e11. doi: 
            10.1016/j.molcel.2023.11.025.
  PUBMED    38134885
REFERENCE   2  (bases 1 to 7279)
  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.molcel.2023.11"; journalName: "Mol Cell"; date: 
            "2023-12-21- 21"; volume: "83"; issue: "24"; pages: "4509-4523.e11"
COMMENT     Sequence Label: MCP-mCherry-TIAL1
FEATURES             Location/Qualifiers
     source          1..7279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1258..1962
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1400..1418)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1668..1687
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3119..3136)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3125..3349
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3395..3823
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3482..3501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3692..3713
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3832..3852)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3837..4166
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4017..4152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4079..4098
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4179..4778
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4179..4198)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4675..4695
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4952..5073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4989..5008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5043..5062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5122..5138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5122..5138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5135..5157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5146..5162
                     /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(5170..5176,5177..5194,5195..5200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5215..5236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5353..5370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5524..6112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5604..5623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6283..7074,7075..7143))
                     /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     6906..6925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7144..7248)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacgggatc gctgaatgga tcagctctaa ctcgcgttca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa aggcgcctgg cgttcgtact taaatatgga actaaccatt
     1141 ccaattttcg ccacgaattc cgactgcgag cttattgtta aggcaatgca aggtctccta
     1201 aaagatggaa acccgattcc ctcagcaatc gcagcaaact ccggcatcta cggatccgtg
     1261 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1321 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1381 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1441 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1501 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1561 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1621 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1681 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1741 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     1801 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1861 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1921 gagggccgcc actccaccgg cggcatggac gagctgtaca agatggaaga cgacgggcag
     1981 ccccggactc tatacgtagg taacctttcc agagatgtga cagaagtcct tatacttcag
     2041 ttgttcagtc agattggacc ctgtaaaagc tgtaaaatga taacagagca tacaagcaat
     2101 gacccatatt gctttgtgga attttatgaa cacagagatg cagctgctgc attagctgct
     2161 atgaatggga gaaaaatttt gggaaaggag gtcaaagtaa actgggcaac cacaccaagt
     2221 agccagaaaa aagatacttc caatcacttc catgtgtttg ttggggattt gagtccagaa
     2281 attacaacag aagatatcaa atcagcattt gccccctttg gtaaaatatc ggatgcccgg
     2341 gtagttaaag acatggcaac tggaaaatcc aaaggctatg gttttgtatc tttttataac
     2401 aaactggatg cagaaaatgc gattgtgcat atgggcggtc agtggttggg tggtcgtcaa
     2461 atccgaacca attgggccac tcgtaaacca cctgcaccta aaagtacaca agaaaacaac
     2521 actaagcagt tgagatttga agatgtagta aaccagtcaa gtccaaaaaa ttgtactgtg
     2581 tactgtggag gaattgcgtc tgggttaaca gatcagctta tgagacagac attctcacca
     2641 tttggacaaa ttatggaaat aagagttttc ccagaaaagg gctattcatt tgtcagattt
     2701 tcaacccatg aaagtgcagc ccatgccatt gtttcggtga acggtactac gattgaagga
     2761 catgtggtta aatgctattg gggtaaagaa tctcctgata tgactaaaaa cttccaacag
     2821 gttgactata gtcaatgggg ccaatggagc caagtgtatg gaaacccaca acagtatgga
     2881 cagtatatgg caaatgggtg gcaagtaccg ccttatggag tatacgggca accatggaat
     2941 caacaaggat ttggagtaga tcaatcacct tctgctgctt ggatgggtgg atttggtgct
     3001 cagcctcccc aaggacaagc tcctccccct gtaatacctc ctcctaacca agccggatat
     3061 ggtatggcaa gttaccaaac acagtgatct agagggcccg tttaaacccg ctgatcagcc
     3121 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3181 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3241 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3301 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3361 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3421 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3481 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3541 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3601 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3661 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3721 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3781 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3841 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3901 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3961 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4021 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4081 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4141 tttttggagg cctaggcttt tgcaaaaagc tcccaaccat gaccgagtac aagcccacgg
     4201 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4261 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4321 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4381 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4441 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4501 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4561 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4621 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4681 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4741 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagc gggactctgg ggttcgaaat
     4801 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4861 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4921 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5041 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5101 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5161 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5221 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5281 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5341 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5401 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5461 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5521 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5581 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5641 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5701 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5761 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5821 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5881 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5941 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6001 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6061 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6121 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6181 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6241 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6301 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6361 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6421 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6481 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6541 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6601 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6661 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6721 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6781 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6841 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6901 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6961 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7021 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7081 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7141 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7201 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7261 aaaagtgcca cctgacgtc
//
