LOCUS       sequence-362473-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Gnat1 protein with N-terminal HA tag from cDNA that 
            contains the full length 3'-UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7537)
  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 7537)
  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 HA-Gnat1 WT-3UTR
FEATURES             Location/Qualifiers
     source          1..7537
                     /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             909..935
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3128..3145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3134..3358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3404..3832
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3491..3510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3701..3722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3841..3861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3846..4175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4026..4161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4088..4107
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4242..5036
                     /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(4296..4315)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4906..4925
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5210..5331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5247..5266)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5301..5320
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5380..5396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5380..5396)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5393..5415)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5404..5420
                     /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(5428..5434,5435..5452,5453..5458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5473..5494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5611..5628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5782..6370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5862..5881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6541..7332,7333..7401))
                     /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     7164..7183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7402..7506)
                     /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 ggaccatgta cccatacgac gtaccagatt acgctggggc tggggccagc gctgaggaga
      961 agcactccag agagctggag aagaagctga aagaggatgc tgagaaggat gcccgcactg
     1021 tgaaactgct gcttctgggt gccggtgaat ccgggaagag cactattgtc aaacagatga
     1081 agattatcca ccaggacggg tattcgctgg aggaatgcct cgagttcatt gccatcatct
     1141 acggcaacac tctgcagtcc atcctggcca tcgttcgggc tatgaccacg ctcaacattc
     1201 agtatggaga ttcagcccgg caggatgatg cccggaagct catgcacatg gcagatacta
     1261 ttgaggaagg cacaatgccc aaggagatgt cagacatcat tcagcgcttg tggaaggact
     1321 cgggtatcca agcttgcttt gaccgagcct cagaatacca gctcaatgac tccgccggct
     1381 actatctctc agacctagag cgtctggtga ctccaggata tgtgcccact gagcaggacg
     1441 tgttgcgttc tcgtgtcaaa accactggta ttatcgagac tcaattctcc ttcaaggacc
     1501 tcaacttcag aatgttcgat gtgggcgggc agcgttccga gcgcaaaaag tggatccact
     1561 gctttgaggg tgtgacgtgc atcattttca tcgctgcgct gagcgcttac gacatggtgc
     1621 tggtggagga cgacgaagtg aaccgaatgc acgagagcct gcacctgttc aacagcatct
     1681 gcaatcatcg ctacttcgcc acaacgtcta tcgtgctctt cctcaacaag aaggacgttt
     1741 tctccgagaa gataaaaaag gcacacctca gcatctgctt ccccgactac gatggaccta
     1801 acacttacga ggatgccggc aactacatca aagtgcagtt cctggagctt aacatgcgac
     1861 gtgatgtgaa ggagatctat tcccacatga cgtgcgctac cgacacacag aacgtcaagt
     1921 ttgtctttga cgctgtcacc gacattatca tcaaggagaa cctcaaagac tgcgggctct
     1981 tctgaggtac ctgaattcag gtgtgtctct tgccctgaga ttctgtagcc ctgcagccct
     2041 gtagccctgt agtcccctca aataccgtcc tttccacaag ctgctgctct tgtagccaca
     2101 cacccaaagg catgaggctt gcagtgcaga ccctccccca agtacccaac tcgaccctgc
     2161 ttttcaccgc ctggctgcac tggcctcttg gacctacagc agccagcctg agctaggaca
     2221 cctcaggact tgggacagct ggaactcaca gtgactcagc agtgccctag tgacccgtct
     2281 gtctccagcc tcttctgcta cccccagcag cagcagcagc tcaccagcag atccccattc
     2341 cacaaacagc cctccctcac cagtttccac gaaggacttg aggtatccct ccctcaagca
     2401 gtaggccggt ccagcctgct acctgcatat gacaagtcca acacgtgtcc acccagtggt
     2461 tagtgactgt ccctcccagt agcctgagga ctccaaagtc caagctctgg tttttgaagc
     2521 ctcgtataga gaagtgagtg actttgacgt gggtgccagg gaaagaaaga cacaagccaa
     2581 acccattccc gaacatcaga ctcagggttt ctccagtccc ttctccctct gggtcttgcc
     2641 cctctgtcct tggacacttc cacaccacac ctcatccacc cacacacaat actcagggct
     2701 ggctggtcaa ggtatgctgt attgaaggag ggtattattg acatagctat gaccaataat
     2761 gaagccatgc tgaggggtgg ctcagagcag agtccaaccc ctattgatca accatgggca
     2821 aagaatatcc catgtgaggt tacaggcaat tccatgtgac catgttgtcc ctcctgattc
     2881 agggcccatt ccttaggagg gagaactgaa aaccaggcct aaaccattag agacaatgtc
     2941 ccccaacccc cagccacttc tacaaccttt gcttaagcct gtggaccaga gccagtgctg
     3001 ctggaagagg caagatgctt ctggacagca agactagggc acaacctctc cctcccttag
     3061 ctagctgcag actcagcaat aaacctttgc accaggtcta gagggcccgt ttaaacccgc
     3121 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3181 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3241 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3301 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3361 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3421 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3481 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3541 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3601 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3661 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3721 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3781 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3841 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3901 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4021 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4081 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4141 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4201 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4261 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4321 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4381 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4441 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4501 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4561 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4621 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4681 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4741 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4801 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4861 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4921 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4981 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5041 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5101 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5161 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5221 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5281 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5341 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5401 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5461 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5521 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5581 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5641 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5701 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5761 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5821 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5881 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5941 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6001 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6061 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6121 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6181 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6241 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6301 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7501 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
