LOCUS       sequence-362449-        6486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repair template for the N-terminal tagging of anillin with 
            mNeonGreen in canine cells using CRISPR/Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  AUTHORS   Husser MC, Ozugergin I, Resta T, Martin VJJ, Piekny AJ
  TITLE     Cytokinetic diversity in mammalian cells is revealed by the 
            characterization of endogenous anillin, Ect2 and RhoA.
  JOURNAL   Open Biol. 2022 Nov;12(11):220247. doi: 10.1098/rsob.220247. Epub 
            2022 Nov 23.
  PUBMED    36416720
REFERENCE   2  (bases 1 to 6486)
  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: "Open 
            Biol."; date: "2022-11"; volume: "12(11):220247. doi"; pages: " 
            10.1098/rsob.220247. Epub 2022 Nov 23"
COMMENT     Sequence Label: pODN-mNG-CfANLN
FEATURES             Location/Qualifiers
     source          1..6486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(2..793,794..862))
                     /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     625..644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(863..967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1035..1053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1359..1378
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1359..1377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2430..3137
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2685..2694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     mobile_element  4609..5376
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     protein_bind    5417..5438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5558..5575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5729..6317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5809..5828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
       61 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
      241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
      301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
      361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
      421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
      481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
      541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
      601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
      661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
      781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
      841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
      901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1021 cattaaccta taaaaatagg cgtatcacga ggccgcccct gcagccgaat tatattattt
     1081 ttgccaaata atttttaaca aaagctctga agtcttcttc atttaaattc ttagatgata
     1141 cttcatctgg aaaattgtcc caattagtag catcacgctg tgagtaagtt ctaaaccatt
     1201 tttttattgt tgtattatct ctaatcttac tactcgatga gttttcggta ttatctctat
     1261 ttttaacttg gagcaggttc cattcattgt ttttttcatc atagtgaata aaatcaactg
     1321 ctttaacact tgtgcctgaa caccatatcc atccggcgta atacgactca ctatagggag
     1381 agcggccgcc agatcttccg gatggctcga gtttttcagc aagattttca gcccaacacg
     1441 tccacaactg ctgcctctac atgccctaac acggacttgc taaaggagaa atgaatcgga
     1501 tacagtaatt gcataaccta tgttgcaatt aattctgagc catccgtaaa gaacggtcgt
     1561 tgagacacag cttgacctgg tggaaagggg gaccctgagc tgagaatggg gtgtttcatc
     1621 ctcttggaca aagggctcct tgtcccaggg tctccgcgtg ttggctgtta agagccaagg
     1681 tggatgcgtg gcttcttcat ccttccaggc ctaacactgt gttggttcgg atgggatcta
     1741 aactaaggag gcagggcggt ttgggtgggg tatgaacatt ccacccgcag cttgctaatg
     1801 tcaagtcgtc tcttcctaga tgggaaaggt ctccagcgcc taggtgggcc actcgccccg
     1861 aatgtccgct tttgggcgac agtccccgtc ggggagggtg ggaagcacgc tgcggacggc
     1921 ccccaggccc gcgggatcgc gaccgcgcgg ccccgcgaac cccccgggga gcgatgacgt
     1981 cgggcgggaa aggtgcaagc accgcgggct gcgagcctaa gctgcggggc tgcgcgggcg
     2041 tcgggtgagg cctcgggccc ggaggcggct gcgccgggga gcgccgcgcc tcccgccgcc
     2101 cgcggcctcc cgagctctgg gtgcgaccca ggggcggcgg cccggacccg gccacgtgac
     2161 ccggggcggg ggcgggggag gggggcgtgg cgggcagagg cgcgtcccgc ccccgcccgg
     2221 cgcgcaggcg cgctgcaccg gcgctgaaat tcaaattcga acggcggcgg gaggctgagt
     2281 ccgacccggc ggctgagggg gcgcgcggca ggagcagttt gggagcgtcg gggtctctcc
     2341 cggcggcgcc cggagccgag actcacttct ttcccttttc cgagcttgga ccggcgtcgt
     2401 cgcgcagtcg agccgcggag tccggggcga tggtgagcaa gggcgaggag gataacatgg
     2461 cctctctccc agcgacacat gagttacaca tctttggctc catcaacggt gtggactttg
     2521 acatggtggg tcagggcacc ggcaatccaa atgatggtta tgaggagtta aacctgaagt
     2581 ccaccaaggg tgacctccag ttctccccct ggattctggt ccctcatatc gggtatggct
     2641 tccatcagta cctgccctac cctgacggga tgtcgccttt ccaggccgcc atggtagatg
     2701 gctccggata ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc cttactgtta
     2761 actaccgcta cacctacgag ggaagccaca tcaaaggaga ggcccaggtg aaggggactg
     2821 gtttccctgc tgacggtcct gtgatgacca actcgctgac cgctgcggac tggtgcaggt
     2881 cgaagaagac ttaccccaac gacaaaacca tcatcagtac ctttaagtgg agttacacca
     2941 ctggaaatgg caagcgctac cggagcactg cgcggaccac ctacaccttt gccaagccaa
     3001 tggcggctaa ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc
     3061 actccaagac cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca
     3121 tggacgagct gtacaaggga ggttcaggag gcagcatgga tccatttacc gaggtgagaa
     3181 agcctgaggg cgcagcctgc catcgccgcc ccgaccccgg ccccggcccg acccgaccct
     3241 tcccaccgcc cgtcgacccc cgggcgcacc gggcggaggg cgcgtgcgcg cgcgtgcgca
     3301 gcgggtgtgc gggggtcggc ccggcgcccc tcccctgccc tcccctgccc tgccctgccc
     3361 ccagccctgc ggcggctcgc gtcggcggct gcggctcagg gaggggggct tgggctgcgg
     3421 cccccgagac ggcagcgcgg agcccccgcg gccccgatcc ggagggtgag gcgcgctgtc
     3481 ctgcggagga ccccgtccct cccggacctg ggcctgctgg cagcgcgagg cgggcagggc
     3541 gcaggatgga gtcgggggct ggaggactcc gcgcaggtgc agccgccgcc gcggagcggg
     3601 cctgcggggg ttcgtgggcc gtggaccgcg gcgggcgagc aggacggccc ggggcgtgac
     3661 ccgagacccg ggatcgagtc cccggccggg ctccctgcac ggagcctgct tctccctctg
     3721 cctgggtctc tgcctctgtg tcgtggataa ataaataaca tctttaaaaa ggaaaaagga
     3781 aatgacagcc aaacgttgac atcagccagt acctgaaacc attgcatccg ttccgataac
     3841 gtcaacagca tctttagagc gggtcgctct gctacatgag gaagaagtcg tcccaacccc
     3901 cgggattgac acgggattga cccaggaacg gggaggaaac atgcgagtcc aacagacacc
     3961 agcaaaatta gtagccaaca atttttcttt ttttttttaa agcagagtaa ggaaatagta
     4021 aacctcttcg tgtgtatgga gcatcacgtt acaagcaata ctctactcgt cttttgtcct
     4081 ttggaaattg aacatggatt cgttacgtca ttgtgtcagt tatttctctc tctccttggc
     4141 cagtcttgtt gagaacatac atctttctag aagatctcct acaatattct cagctgccat
     4201 ggaaaatcga tgttcttctt ttattctctc aagattttca ggctgtatat taaaacttat
     4261 attaagaact atgctaacca cctcatcagg aaccgttgta ggtggcgtgg gttttcttgg
     4321 caatcgactc tcatgaaaac tacgagctaa atattcaata tgttcctctt gaccaacttt
     4381 attctgcatt ttttttgaac gaggtttaga gcaagcttca ggaaactgag acaggaattt
     4441 tattaaaaat ttaaattttg aagaaagttc agggttaata gcatccattt tttgctttgc
     4501 aagttcctca gcattcttaa caaaagacgt ctcttttgac atgtttaaag tttaaacctc
     4561 ctgtgtgaaa ttattatccg ctcataattc cacacattat acgagccggg tgatgctgcc
     4621 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat
     4681 cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     4741 acatcagcgc tatctctgct ctcactgccg taaaacatgg caactgcagt tcacttacac
     4801 cgcttctcaa cccggtacgc accagaaaat cattgatatg gccatgaatg gcgttggatg
     4861 ccgggcaaca gcccgcatta tgggcgttgg cctcaacacg attttacgtc acttaaaaaa
     4921 ctcaggccgc agtcggtaac ctcgcgcata cagccgggca gtgacgtcat cgtctgcgcg
     4981 gaaatggacg aacagtgggg ctatgtcggg gctaaatcgc gccagcgctg gctgttttac
     5041 gcgtatgaca gtctccggaa gacggttgtt gcgcacgtat tcggtgaacg cactatggcg
     5101 acgctggggc gtcttatgag cctgctgtca ccctttgacg tggtgatatg gatgacggat
     5161 ggctggccgc tgtatgaatc ccgcctgaag ggaaagctgc acgtaatcag caagcgatat
     5221 acgcagcgaa ttgagcggca taacctgaat ctgaggcagc acctggcacg gctgggacgg
     5281 aagtcgctgt cgttctcaaa atcggtggag ctgcatgaca aagtcatcgg gcattatctg
     5341 aacataaaac actatcaata agttggagtc attacctacg agccggaagc ataaagtgta
     5401 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgccaa
     5461 ttgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5521 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5581 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5641 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5701 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5761 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5821 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5881 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5941 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6001 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6061 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6121 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6181 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6241 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6361 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6421 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6481 ctgaca
//
