LOCUS       sequence-236352-        2210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly DE part for gV1 gRNA (guide RNA for S. 
            pyogenes Cas9; sequence from doi: [10.15252/msb.20145735]). 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: UC17m
FEATURES             Location/Qualifiers
     source          1..2210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..23
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(27..70)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    73..94
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_RNA        133..208
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    226..246
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(252..269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(423..1011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(503..522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1182..1973,1974..2042))
                     /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     1805..1824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2043..2147)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
       61 ggatgatttc tggaattcgc ggccgcttct agagactagt gggtctcata cttttacacc
      121 tagctcagtc ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
      181 acttgaaaaa gtggcaccga gtcggtgctt ttttaggtag agacctacta gtagcggccg
      241 ctgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      301 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      361 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      421 tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      481 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      541 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      601 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      661 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      721 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      781 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      841 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      901 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      961 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1021 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1081 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1141 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1201 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1261 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1321 cgcgacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1381 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1441 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1501 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1561 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1621 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1681 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1741 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1801 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1861 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1921 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1981 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2041 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2101 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2161 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
//
