LOCUS       sequence-454857-        2115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for PCR amplification for gRNA cloning plasmid. The PCR 
            product recombines in vivo with a PCR product from pSpec5', to clone
            gRNA plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2115)
  TITLE     Engineering Adaptive Alleles for Escherichia coli Growth on Sucrose 
            Using the EasyGuide CRISPR System
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2115)
  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            "Unpublished"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: pSpec3'
FEATURES             Location/Qualifiers
     source          1..2115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(36..104,105..896)
                     /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(254..273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1527..2115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2016..2035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgacagcta gctcagtcct aggtataata ctagtatgag tattcaacat ttccgtgtcg
       61 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      121 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      181 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      241 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      301 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      361 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      421 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      481 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      541 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      601 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      661 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      721 ttgctgataa atctggagcc ggtgagcgtg ggagccgcgg tatcattgca gcactggggc
      781 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      841 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      901 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      961 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttccctt aacgctatgg
     1081 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     1141 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     1201 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     1261 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     1321 aaaggcgaga tcaccaaggt agtcggcaaa taagatgccg ctcgccagtc gattggctga
     1381 gctcataagt tcctattccg aagttccgcg aacgcgtaaa ggatctaggt gaagatcctt
     1441 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1501 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1561 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1621 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     1681 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1741 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1801 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1861 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1921 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1981 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2041 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2101 cggagcctat ggaaa
//
