LOCUS       sequence-326579-        2526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA synthesis vector for smyhc1_977 (zebrafish slow myosin heavy 
            chain 1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2526)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 2526)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: pT7gRNA-smyhc1_977
FEATURES             Location/Qualifiers
     source          1..2526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      163..751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     652..671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     905..922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1039..1060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1075..1080,1081..1098,1099..1105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1113..1129
                     /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)."
     primer_bind     1118..1140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1137..1153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1137..1153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1177..1195
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        1214..1289
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1317..1334)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1317..1333)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1326..1348)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1553..1657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1658..1726,1727..2518)
                     /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     complement(1876..1895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
       61 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
      121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      781 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      841 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      901 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      961 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1021 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     1081 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     1141 aaacagctat gaccatgatt acgccaagct tgaaattaat acgactcact ataggcagtg
     1201 gcaatcaact cttgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     1261 aacttgaaaa agtggcaccg agtcggtgcg gatccccggg taccgagctc gaattcactg
     1321 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1381 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1441 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     1501 catctgtgcg gtatttcaca ccgcataagg tggcactttt cggggaaatg tgcgcggaac
     1561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2521 gtcaga
//
