LOCUS       sequence-418358-        3150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for long single strand DNA donnor preparation for Twin Strep tag 
            knock-in at C ter of human RPL3. Digest with nt.BspQI and nb.BsrDI 
            to get ssDNA. This plasmid will cause re-edit and fail knock-in..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3150)
  TITLE     RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3150)
  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            "Unpublished"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: pUC-ODN-hsRPL3-CterTwStrep
FEATURES             Location/Qualifiers
     source          1..3150
                     /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"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(620..625,626..649,650..685,686..709)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(929..945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(929..945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(942..964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    953..969
                     /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(977..983,984..1001,1002..1007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1022..1043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1160..1177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1331..1919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1411..1430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2090..2881,2882..2950))
                     /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     2713..2732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2951..3055)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3123..3141
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     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 ccagtgaatt cgagctcggt acccggggct
      421 cttcccgggg taggggtggt cttgatgcct taggcctctc ttgtggctgt ggagactggc
      481 ccagagtctt ggtctcctct agagtgggca tctgctgtca cctgtcatct tctacctctg
      541 accactgcct ctctccccac agggaccact caaaaaggac cgaattgcaa aggaagaagg
      601 agctaagctt tccggaggga gcgcttggag ccacccgcag ttcgaaaaag gtggaggttc
      661 tggcggtgga tcgggaggtt cagcgtggtc tcacccccaa tttgagaagt aatgccagga
      721 acagattttg cagttggtgg ggtctcaata aaagttattt tccactgaca tcttgcctct
      781 ttcttcagga gagaggctgg gactggggag gcagatgggg gcccttgagg tgccttgtgc
      841 tccccaaggc cactgccctc ctttctgtag cttcgctggc attgcctcta gagtcgacct
      901 gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      961 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1021 aatgagtgag ctaactcaca ttaattgcgt tgcgctcaca gcccgctttc cagtcgggaa
     1081 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1141 ttgggcgctg ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1501 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1741 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2101 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2221 taataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2401 tagctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2461 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2521 tcggtcctcc gatcgttgtc agaagtaagt tggccgctgt gttatcactc atggttatgg
     2581 cagcactaca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2701 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3001 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3061 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3121 aaaataggcg tatcacgagg ccctttcgtc
//
