LOCUS       sequence-454859-        2052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for PCR amplification for gRNA cloning. The PCR product 
            recombines in vivo with a PCRs products from pOliSpec5' and 
            oligo-spacer amplifications, to clone gRNA plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2052)
  TITLE     Engineering Adaptive Alleles for Escherichia coli Growth on Sucrose 
            Using the EasyGuide CRISPR System
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2052)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pOliSpec3'
FEATURES             Location/Qualifiers
     source          1..2052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(37..105,106..897)
                     /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(255..274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1464..2052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1953..1972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgacagcta gctcagtcct aggtataata ggatccatga gtattcaaca tttccgtgtc
       61 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      121 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      181 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      241 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
      301 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      361 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
      421 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      481 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
      541 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
      601 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      661 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      721 attgctgata aatctggagc cggtgagcgt gggagccgcg gtatcattgc agcactgggg
      781 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      841 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
      901 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      961 aggatctagg tgaagatcct ttttgataat ctcatggatc ctcccttaac gctatggaac
     1021 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
     1081 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
     1141 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
     1201 gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
     1261 ggcgagatca ccaaggtagt cggcaaataa gatgccgctc gccagtcgat tggctgagct
     1321 cataagttcc tattccgaag ttccgcgaac gcgtaaagga tctaggtgaa gatccttttt
     1381 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1441 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1501 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1561 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1621 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1681 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1741 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1801 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1861 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1921 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1981 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2041 agcctatgga aa
//
