LOCUS       sequence-91105-e        3345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains multiple cloning sites (MCSs) for cloning and transient 
            expression of single or multiple guide RNA (gRNA) expression 
            cassettes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3345)
  TITLE     Sheen lab CRISPR plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3345)
  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            "Unpublished"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: pUC119-MCS
FEATURES             Location/Qualifiers
     source          1..3345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(474..491)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(474..490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(483..505)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      703..1158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(790..809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1000..1021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1175..1194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1294..1316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1354..1372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1440..1544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1545..1613,1614..2405)
                     /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(1763..1782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2576..3164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3065..3084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3318..3335
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgggcgc gccaagctta
      241 ggcctgtcga cgcccgggcg gtaccgcgat cgctcgcgac ctgcaggcat aaagccgtca
      301 gtgtccgcat aaagaaccac ccataatacc cataatagct gtttgccatc gctaccttag
      361 gaccgttata gttaattacc ctgttatccc tattaattaa gagctcgcta ccttaagaga
      421 ggatatcggc gcgcctctag aatttaaatg gatcctacgt actcgaggaa ttcactggcc
      481 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
      541 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
      601 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
      661 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
      721 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      781 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      841 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      901 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      961 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1021 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     1081 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1141 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1201 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     1261 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1321 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1381 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2701 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     2761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3181 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3241 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3301 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
