LOCUS       sequence-322310-        2915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-targeted-plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2915)
  TITLE     Dongchang Sun Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2915)
  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            "Unpublished"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: pCR1
FEATURES             Location/Qualifiers
     source          1..2915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      760..1348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1249..1268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1502..1519
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1636..1657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1672..1677,1678..1695,1696..1702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1710..1726
                     /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     1715..1737
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1734..1750
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1734..1750
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2049..2066)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2049..2065)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2058..2080)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2274..2293)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2393..2415
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2453..2471)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2539..2643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2644..2712,2713..2915,1..589)
                     /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(2862..2881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
       61 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      121 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      181 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      241 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
      301 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      361 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      421 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      481 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      541 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
      601 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
      661 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
      721 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      781 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      841 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      901 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      961 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1021 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1081 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1141 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1201 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1261 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1321 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1381 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1441 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1501 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1561 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1621 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     1681 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1741 cagctatgac catgattacg ccaagcttct ttttatcccc gctggcgcgg ggaactcgcg
     1801 accgctcaga aattccagac ccgatccaaa ctttttatcc ccgctggcgc ggggaactct
     1861 caacattatc aattacaacc gacagggagc cctttttatc cccgctggcg cggggaactc
     1921 agcgtgttcg gcatcacctt tggcttcggc tgctttttat ccccgctggc gcggggaact
     1981 ctgcgtgagc gtatcgccgc gcgtctgcga aagcttttta tccccgctgg cgcggggaac
     2041 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2101 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2161 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     2221 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     2281 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2341 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2401 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2461 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2521 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2581 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2641 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2701 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2761 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2821 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2881 tcccgtattg acgccgggca agagcaactc ggtcg
//
