LOCUS       sequence-225357-        2149 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 2149)
  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 2149)
  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_T2_AmpR
FEATURES             Location/Qualifiers
     source          1..2149
                     /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(842..1633,1634..1702))
                     /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     1465..1484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1703..1807)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(1910..2149,1..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 gttaaaataa ggctagtccg ttatcagccg gcttatcggt
      601 cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag
      661 aaagatgaat gactgtccac gacgctatac ccaaaagaaa aagcttgaaa gaatcgatcg
      721 atcccgaatg agtagtacag cggccgcgat tatcaaaaag gatcttcacc tagatccttt
      781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      961 ccagtgctgc aatgataccg cgggacccac gctcaccggc tccagattta tcagcaataa
     1021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1801 ttccgcgcac atttccccga aaagtgccac ctgtcatgac caaaatccct taacgtgagt
     1861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgc
//
