LOCUS       sequence-225359-        2130 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 2130)
  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 2130)
  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_T3_AmpR
FEATURES             Location/Qualifiers
     source          1..2130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     250..269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        484..518
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(join(823..1614,1615..1683))
                     /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     1446..1465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1684..1788)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(1891..2130,1..349)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
       61 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      421 gttatcccct gattctgtgg ataaccgtgc ggcccgagag catcgatcct agcatggatg
      481 catttgacag ctagctcagt cctaggtata atgctagcgg taaaattgtc aacacgcagt
      541 tttagagcta gaaatagcaa gttaaaatcc ggcttatcgg tcagtttcac ctgatttacg
      601 taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa tgactgtcca
      661 cgacgctata cccaaaagaa aaagcttgaa agaatcgatc gatcccgaat gagtagtaca
      721 gcggccgcga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      961 gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1801 aaaagtgcca cctgtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1861 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1921 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1981 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2041 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2101 cgctctgcta atcctgttac cagtggctgc
//
