LOCUS       sequence-362446-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Msi1 clone with N-terminal flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  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 6501)
  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 flag-Msi1
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..50)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     241..262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(381..401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        386..715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      566..701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     628..647
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             782..1576
                     /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(836..855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1446..1465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1750..1871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1787..1806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1841..1860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1920..1936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1920..1936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1933..1955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1944..1960
                     /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(1968..1974,1975..1992,1993..1998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2013..2034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2151..2168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2322..2910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2402..2421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3081..3872,3873..3941))
                     /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     3704..3723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3942..4046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4121..4140)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4312..4691
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4692..4895
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4846..4866
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4940..4959
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4940..4958
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5019..5042
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      5857..5866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6169..6186)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6175..6399
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6445..6501,1..372)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
       61 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      121 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      181 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      241 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      301 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      361 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      541 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
      661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      721 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
      781 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      841 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      901 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      961 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     1021 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     1081 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     1141 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     1201 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     1261 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     1321 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     1381 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     1441 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     1501 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     1561 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     1621 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1681 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1741 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1801 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1861 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     1921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     4081 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     4141 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     4201 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     4261 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4321 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4381 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4441 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4501 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4561 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4621 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4681 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4741 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4801 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4861 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     4921 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
     4981 taaacttaag cttggtaccg agctcggatc ccaccatgga ttacaaggat gacgacgata
     5041 agatggagac tgacgcgccc cagcccggcc tcgcctcccc ggactcgccg cacgacccct
     5101 gcaagatgtt catcggagga ctcagttggc agaccacgca ggaagggctg cgcgaatact
     5161 tcggccagtt cggggaggtg aaagagtgtc tggtgatgcg ggaccccctg accaaaagat
     5221 ccaggggttt cggcttcgtc actttcatgg accaggcggg ggtggataaa gtgctggcgc
     5281 aatcgcggca cgagctcgac tccaaaacaa ttgaccccaa ggtggccttt cctcgaagag
     5341 cacagcctaa gatggtcact cggacgaaga agatcttcgt gggggggctg tctgtgaaca
     5401 ccacggtgga agatgtgaaa cactatttcg agcagttcgg aaaggtggat gatgccatgc
     5461 tgatgttcga caaaaccacc aacaggcaca gagggtttgg atttgtcacg tttgagagcg
     5521 aggacatcgt agagaaagtt tgtgagatcc acttccatga aatcaacaac aaaatggtgg
     5581 aatgcaagaa agcccagcca aaggaggtga tgtccccgac aggctcagcc cggggcaggt
     5641 ctcgggtcat gccctacgga atggatgcct tcatgctggg tattgggatg ctgggttacc
     5701 cagggttcca agccacgacc tacgccagcc ggagttacac aggccttgcc cctggttaca
     5761 cctaccagtt ccccgaattc cgtgtagagc ggagccctct cccgagcgcc ccagtcctcc
     5821 ccgagctcac agctatccct ctcacggctt atgggcccat ggcggcggca gcggcggcgg
     5881 cagctgtagt tcgagggaca ggctctcacc cctggacgat ggctccccct ccaggttcca
     5941 ctcccagccg cacagggggc ttcctaggga ccacaagccc cggccccatg gctgagctct
     6001 acggggcagc caaccaggac tccggggtca gcagttacat cagcgccgcc agccccgccc
     6061 ccagcactgg tttcggccac agtcttgggg gtcccttgat tgccacagcc ttcaccaatg
     6121 ggtaccactg actcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     6181 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     6241 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     6301 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     6361 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     6421 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6481 tggtggttac gcgcagcgtg a
//
