LOCUS       sequence-245418-        2210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly DE part for gQi gRNA (guide RNA for S. 
            pyogenes Cas9; sequence from DOI: 10.1016/j.cell.2013.02.022). 
            Please see Supplemental Documents for annotated Genbank file..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2210)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2210)
  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"
COMMENT     Sequence Label: UCT1m
FEATURES             Location/Qualifiers
     source          1..2210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..67
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(71..114)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    117..138
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_RNA        177..252
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    270..290
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(296..313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(467..1055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(547..566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1226..2017,2018..2086))
                     /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     1849..1868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2087..2191)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
       61 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat
      121 tcgcggccgc ttctagagac tagtgggtct catactaact ttcagtttag cggtctgttt
      181 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
      241 ccgagtcggt gcttttttgc ttagagacct actagtagcg gccgctgcag gcttcctcgc
      301 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      361 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc
      481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      901 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgcgac ccacgctcac
     1381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
//
