LOCUS       sequence-487893-        3025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (C' terminus) for C>T base editing, 3xHA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3025)
  TITLE     Fragmid
REFERENCE   2  (bases 1 to 3025)
  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"
COMMENT     Sequence Label: AA557
FEATURES             Location/Qualifiers
     source          1..3025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             468..716
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             729..776
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             783..863
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(935..951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(935..951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(948..970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    959..975
                     /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)."
     promoter        complement(join(983..989,990..1007,1008..1013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1028..1049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1166..1183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1337..1925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1417..1436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2103..2912)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2826..2845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2904..2923
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacggg tccgggagcg gcgggagcgg ggggagcact aatctgagcg
      481 acatcattga gaaggagact gggaaacagc tggtcattca ggagtccatc ctgatgctgc
      541 ctgaggaggt ggaggaagtg atcggcaaca agccagagtc tgacatcctg gtgcacaccg
      601 cctacgacga gtccacagat gagaatgtga tgctgctgac ctctgacgcc cccgagtata
      661 agccttgggc cctggtcatc caggattcta acggcgagaa taagatcaag atgctgagcg
      721 gaggctccaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagggat
      781 cctacccata cgatgttcca gattacgctt atccctacga cgtgcctgat tatgcatacc
      841 catatgatgt ccccgactat gccggaagca cgtcttcatc ggatcccggg cccgtcgact
      901 gcagaggcct gcatgcaagc ttggtgtaat catggtcata gctgtttcct gtgtgaaatt
      961 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1021 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1081 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1141 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1741 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2041 aaaaatgaag ttttaaatca agcccaatct gaataatgtt acaaccaatt aaccaattct
     2101 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2161 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2221 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2281 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2341 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2401 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2461 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2521 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2581 tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca
     2641 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2701 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2761 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     2821 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2881 ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     2941 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     3001 ttttgagaca cgggccagag ctgca
//
