LOCUS       sequence-382818-        4632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown mouse KIF5A, KIF5B and KIF5C simultaneously. 
            mScarlet::miRNA(KIF5C)::miRNA(KIF5B)::miRNA(KIF5A)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4632)
  AUTHORS   Niwa S, Chiba K
  TITLE     Generation of recombinant and chickenized scFv versions of an 
            anti-kinesin monoclonal antibody H2.
  JOURNAL   Cytoskeleton (Hoboken). 2023 Apr 10. doi: 10.1002/cm.21756.
  PUBMED    37036074
REFERENCE   2  (bases 1 to 4632)
  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: 
            "Cytoskeleton (Hoboken). 2023 Apr 10. doi: 10.1002/cm.21756."
COMMENT     Sequence Label: pSN879
FEATURES             Location/Qualifiers
     source          1..4632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        373..752
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        753..956
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     907..927
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1001..1019
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1049..1073
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1082..1777
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1215..1233)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1483..1502
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     ncRNA           1840..1866
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1930..1971
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1978..2004
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           2068..2109
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           2116..2142
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           2206..2247
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(2276..2300)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2342..2361)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2386..2434
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(2605..2623)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2628..2644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2775..2792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2885..3473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2965..2984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3644..4435,4436..4504))
                     /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     4267..4286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4505..4609)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccgcatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      421 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      481 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      541 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      601 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      661 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      721 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      781 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      841 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      901 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
      961 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     1021 agtcccaagc tggctagtta agctatcaac aagtttgtac aaaaaagcag gctttaaaac
     1081 catggtgagc aagggcgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga
     1141 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1201 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga
     1261 catcctgtcc cctcagttca tgtacggctc cagggccttc accaagcacc ccgccgacat
     1321 ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1381 cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat
     1441 ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa
     1501 gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg
     1561 cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg acttcaagac
     1621 cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt
     1681 ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg
     1741 ccgccactcc accggcggca tggacgagct gtacaagtaa gctaagcact tcgtggccgt
     1801 cgatcgttta aagggaggta gtgagtcgac cagtggatcc tggaggcttg ctgaaggctg
     1861 tatgctgttc ttgtgacttc tggtcgtagt tttggccact gactgactac gaccaagtca
     1921 caagaacagg acacaaggcc tgttactagc actcacatgg aacaaatggc ccagatcctg
     1981 gaggcttgct gaaggctgta tgctgttcat ttgcagtctg aaccttgttt tggccactga
     2041 ctgacaaggt tcactgcaaa tgaacaggac acaaggcctg ttactagcac tcacatggaa
     2101 caaatggccc agatcctgga ggcttgctga aggctgtatg ctgaaatcgg caaagcacct
     2161 tgatgttttg gccactgact gacatcaagg ttttgccgat ttcaggacac aaggcctgtt
     2221 actagcactc acatggaaca aatggcccag atctggccgc actcgagata tctagaccca
     2281 gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgctgatgg gggaggctaa
     2341 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     2401 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     2461 tggcactctg tcgatacccc accgtgaccc cattggggcc aatacgcccg cgtttcttcc
     2521 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     2581 gcggcaggcc ctgccatagc atcccctata gtgagtcgta ttacatggtc atagctgttt
     2641 cctggcagct ctggcccgtg ctgggcctca tgggccttcc gctcactgcc cgctttccag
     2701 tcgggaaacc tgtcgtgcca gctgcattaa catggtcata gctgtttcct tgcgtattgg
     2761 gcgctctccg cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg
     2821 gtgcctaatg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3481 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3541 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3601 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3661 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3721 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3781 cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3841 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3901 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3961 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4021 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4081 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4141 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4201 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4261 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4321 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4381 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4441 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4501 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4561 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4621 gaaaagtgcc ac
//
