LOCUS       sequence-229450-        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"
     promoter        55..89
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(join(394..1185,1186..1254))
                     /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     1017..1036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1255..1359)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1462..2050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1951..1970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgattctgtg gataaccgtg cggcccgaga gcatcgatcc tagcatggat gcatttgaca
       61 gctagctcag tcctaggtat aatgctagcg gtaaaattgt caacacgcag ttttagagct
      121 agaaatagca agttaaaatc cggcttatcg gtcagtttca cctgatttac gtaaaaaccc
      181 gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgctat
      241 acccaaaaga aaaagcttga aagaatcgat cgatcccgaa tgagtagtac agcggccgcg
      301 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      361 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      421 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      481 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc
      541 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      601 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      661 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      721 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      781 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      841 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
      901 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
      961 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1021 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1081 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1141 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1201 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1321 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1381 acctgtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1441 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1501 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1561 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1621 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1681 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1741 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1801 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1861 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1921 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1981 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2041 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2101 tgctcacatg ttctttcctg cgttatcccc
//
