LOCUS       sequence-476554-        2584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  guide for aeBlue.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2584)
  AUTHORS   Collins M, Lau MB, Ma W, Shen A, Wang B, Cai S, La Russa M, Jewett 
            MC, Qi LS
  TITLE     A frugal CRISPR kit for equitable and accessible education in gene 
            editing and synthetic biology.
  JOURNAL   Nat Commun. 2024 Aug 3;15(1):6563. doi: 10.1038/s41467-024-50767-2.
  PUBMED    39095367
REFERENCE   2  (bases 1 to 2584)
  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: "2024-08-3"; volume: "15(1):6563. doi"; pages: " 
            10.1038/s41467-024-50767-2"
COMMENT     Sequence Label: pSLQ14103 aeBlue-targeting_sgRNA
FEATURES             Location/Qualifiers
     source          1..2584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(813..859)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1012..1029
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(1085..1102)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1118..1147)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_RNA        complement(1168..1243)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(1264..1298)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     terminator      1316..1359
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1363..1381)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1449..1553
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1554..1622,1623..2414)
                     /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(1772..1791)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctgtggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcctgca gtctagactc
      781 gagtaaggat ccagttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
      841 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg agaccccaca
      901 ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc
      961 gcgctactgc cgccaggcaa attctgtttt atcagaccgc ttctgcgttc tgatttaatc
     1021 tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc caagctggag accgtttaaa
     1081 ctcaatgatg atgatgatga tggtcgacgg cgctattcag atcctcttct gagatgagtt
     1141 tttgttcggg cccaagcttc aaaaaaagca ccgactcggt gccacttttt caagttgata
     1201 acggactagc cttattttaa cttgctattt ctagctctaa aacggtggaa aagatacgca
     1261 tcaactagta ttatacctag gactgagcta gctgtcaaag atctttagaa ttccagaaat
     1321 catccttagc gaaagctaag gatttttttt atctgaaatt ctgcctcgtg atacgcctat
     1381 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1441 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1501 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1561 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1621 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1681 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1741 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1801 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1861 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1921 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1981 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2041 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2101 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2161 aacaattaat agactggatg gaggcggata aagttgcggg accacttctg cgctcggccc
     2221 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2281 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2341 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2401 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2461 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2521 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2581 cttc
//
