LOCUS       sequence-362429-        7540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Gnat1 protein with N-terminal T7 tag from cDNA that 
            contains the mutant 3'-UTR where the TAG binding sites for MSI1 are 
            mutated to TGA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7540)
  AUTHORS   Matalkah F, Jeong B, Sheridan M, Horstick E, Ramamurthy V, Stoilov P
  TITLE     The Musashi proteins direct post-transcriptional control of protein 
            expression and alternate exon splicing in vertebrate photoreceptors.
  JOURNAL   Commun Biol. 2022 Sep 24;5(1):1011. doi: 10.1038/s42003-022-03990-w.
  PUBMED    36153373
REFERENCE   2  (bases 1 to 7540)
  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: "Commun 
            Biol."; date: "2022-09-24"; volume: "5(1):1011. doi"; pages: " 
            10.1038/s42003-022-03990-w"
COMMENT     Sequence Label: pcDNA T7-Gnat1 Mut-3UTR
FEATURES             Location/Qualifiers
     source          1..7540
                     /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             906..938
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     primer_bind     complement(3131..3148)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3137..3361
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3407..3835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3494..3513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3704..3725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3844..3864)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3849..4178
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4029..4164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4091..4110
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4245..5039
                     /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(4299..4318)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4909..4928
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5213..5334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5250..5269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5304..5323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5383..5399)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5383..5399)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5396..5418)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5407..5423
                     /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(5431..5437,5438..5455,5456..5461))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5476..5497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5614..5631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5785..6373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5865..5884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6544..7335,7336..7404))
                     /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     7167..7186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7405..7509)
                     /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 gctggtgctg
      901 ggaccatggc tagcatgact ggtggacagc aaatgggtgg ggctggggcc agcgctgagg
      961 agaagcactc cagagagctg gagaagaagc tgaaagagga tgctgagaag gatgcccgca
     1021 ctgtgaaact gctgcttctg ggtgccggtg aatccgggaa gagcactatt gtcaaacaga
     1081 tgaagattat ccaccaggac gggtattcgc tggaggaatg cctcgagttc attgccatca
     1141 tctacggcaa cactctgcag tccatcctgg ccatcgttcg ggctatgacc acgctcaaca
     1201 ttcagtatgg agattcagcc cggcaggatg atgcccggaa gctcatgcac atggcagata
     1261 ctattgagga aggcacaatg cccaaggaga tgtcagacat cattcagcgc ttgtggaagg
     1321 actcgggtat ccaagcttgc tttgaccgag cctcagaata ccagctcaat gactccgccg
     1381 gctactatct ctcagaccta gagcgtctgg tgactccagg atatgtgccc actgagcagg
     1441 acgtgttgcg ttctcgtgtc aaaaccactg gtattatcga gactcaattc tccttcaagg
     1501 acctcaactt cagaatgttc gatgtgggcg ggcagcgttc cgagcgcaaa aagtggatcc
     1561 actgctttga gggtgtgacg tgcatcattt tcatcgctgc gctgagcgct tacgacatgg
     1621 tgctggtgga ggacgacgaa gtgaaccgaa tgcacgagag cctgcacctg ttcaacagca
     1681 tctgcaatca tcgctacttc gccacaacgt ctatcgtgct cttcctcaac aagaaggacg
     1741 ttttctccga gaagataaaa aaggcacacc tcagcatctg cttccccgac tacgatggac
     1801 ctaacactta cgaggatgcc ggcaactaca tcaaagtgca gttcctggag cttaacatgc
     1861 gacgtgatgt gaaggagatc tattcccaca tgacgtgcgc taccgacaca cagaacgtca
     1921 agtttgtctt tgacgctgtc accgacatta tcatcaagga gaacctcaaa gactgcgggc
     1981 tcttctgagg tacctgaatt caggtgtgtc tcttgccctg agattctgtg accctgcagc
     2041 cctgtgaccc tgtgatcccc tcaaataccg tcctttccac aagctgctgc tcttgtgacc
     2101 acacacccaa aggcatgagg cttgcagtgc agaccctccc ccaagtaccc aactcgaccc
     2161 tgcttttcac cgcctggctg cactggcctc ttggacctac agcagccagc ctgagctgag
     2221 acacctcagg acttgggaca gctggaactc acagtgactc agcagtgccc tgatgacccg
     2281 tctgtctcca gcctcttctg ctacccccag cagcagcagc agctcaccag cagatcccca
     2341 ttccacaaac agccctccct caccagtttc cacgaaggac ttgaggtatc cctccctcaa
     2401 gcagtgagcc ggtccagcct gctacctgca tatgacaagt ccaacacgtg tccacccagt
     2461 ggttgatgac tgtccctccc agtgacctga ggactccaaa gtccaagctc tggtttttga
     2521 agcctcgtat gaagaagtga gtgactttga cgtgggtgcc agggaaagaa agacacaagc
     2581 caaacccatt cccgaacatc agactcaggg tttctccagt cccttctccc tctgggtctt
     2641 gcccctctgt ccttggacac ttccacacca cacctcatcc acccacacac aatactcagg
     2701 gctggctggt caaggtatgc tgtattgaag gagggtatta ttgacatgac tatgaccaat
     2761 aatgaagcca tgctgagggg tggctcagag cagagtccaa cccctattga tcaaccatgg
     2821 gcaaagaata tcccatgtga ggttacaggc aattccatgt gaccatgttg tccctcctga
     2881 ttcagggccc attccttgag agggagaact gaaaaccagg cctaaaccat tgaagacaat
     2941 gtcccccaac ccccagccac ttctacaacc tttgcttaag cctgtggacc agagccagtg
     3001 ctgctggaag aggcaagatg cttctggaca gcaagactga ggcacaacct ctccctccct
     3061 tgactgactg cagactcagc aataaacctt tgcaccaggt ctagagggcc cgtttaaacc
     3121 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3181 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3241 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3301 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3361 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3421 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3481 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3541 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3601 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3661 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3721 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3781 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3841 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3901 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4021 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4081 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4141 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4201 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4261 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4321 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4381 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4441 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4501 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4561 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4621 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4681 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4741 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4801 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4861 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4921 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4981 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5041 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5101 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5161 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5221 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5281 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5341 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5401 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5461 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5521 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5581 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6661 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6841 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7141 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7441 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7501 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
