LOCUS       sequence-229449-        2176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes sgRNA targeting position 6 in pColE1_70a_deGFP_KanR, 
            truncated in the 3' end.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2176)
  AUTHORS   Collias D, Marshall R, Collins SP, Beisel CL, Noireaux V
  TITLE     An educational module to explore CRISPR technologies with a 
            cell-free transcription-translation system.
  JOURNAL   Synth Biol (Oxf). 2019 Jan 21;4(1):ysz005. doi: 
            10.1093/synbio/ysz005. eCollection 2019.
  PUBMED    32995532
REFERENCE   2  (bases 1 to 2176)
  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: "Synth Biol 
            (Oxf)."; date: "2019-01-21"; volume: "4(1):ysz005. doi"; pages: " 
            10.1093/synbio/ysz005. eCollection 2019"
COMMENT     Sequence Label: pColE1_sgRNA_T1_AmpR
FEATURES             Location/Qualifiers
     source          1..2176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..75
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        96..171
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(426..1217,1218..1286))
                     /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     1049..1068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1287..1391)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1494..2082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1983..2002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtgcggc ccgagagcat cgatcctagc atggatgcat ttgacagcta gctcagtcct
       61 aggtataatg ctagcggtaa aattgtcaac acgcagtttt agagctagaa atagcaagtt
      121 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cccggcttat
      181 cggtcagttt cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg
      241 gcagaaagat gaatgactgt ccacgacgct atacccaaaa gaaaaagctt gaaagaatcg
      301 atcgatcccg aatgagtagt acagcggccg cgattatcaa aaaggatctt cacctagatc
      361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
      421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
      481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
      541 ggccccagtg ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca
      601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
      661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
      721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
      781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
      841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
      901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
      961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     1021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     1081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     1141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     1201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     1261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     1321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     1381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgtca tgaccaaaat cccttaacgt
     1441 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1561 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1621 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1681 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1741 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1801 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1861 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1921 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1981 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2041 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2101 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2161 cctgattctg tggata
//
