LOCUS       sequence-236353-        2210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly DE part for gN2 gRNA (guide RNA for S. 
            pyogenes Cas9; sequence designed with NUPACK for stable tracrRNA 
            structure and open targeting region)..
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: UC20m
FEATURES             Location/Qualifiers
     source          1..2210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..24
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(28..71)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    74..95
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_RNA        134..209
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    227..247
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(253..270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(424..1012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(504..523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1183..1974,1975..2043))
                     /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     1806..1825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2044..2148)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta
       61 aggatgattt ctggaattcg cggccgcttc tagagactag tgggtctcat actctcctcc
      121 tcccccccaa tatgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      181 aacttgaaaa agtggcaccg agtcggtgct tttttaggta gagacctact agtagcggcc
      241 gctgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      301 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      361 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      421 gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      601 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      841 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1321 gcgcgaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2161 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
//
