LOCUS       sequence-392215-        4930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert TriTag (mTagBFP) into the C-terminus 
            of human SEC61B gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4930)
  AUTHORS   Liang Y, Xu H, Cheng T, Fu Y, Huang H, Qian W, Wang J, Zhou Y, Qian 
            P, Yin Y, Xu P, Zou W, Chen B
  TITLE     Gene activation guided by nascent RNA-bound transcription factors.
  JOURNAL   Nat Commun. 2022 Nov 28;13(1):7329. doi: 10.1038/s41467-022-35041-7.
  PUBMED    36443367
REFERENCE   2  (bases 1 to 4930)
  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: "Nat 
            Commun."; date: "2022-11-28"; volume: "13(1):7329. doi"; pages: " 
            10.1038/s41467-022-35041-7"
COMMENT     Sequence Label: pCA22-SEC61B 5'HA-TriTag (mTagBFP)-3'HA (HDR d...
FEATURES             Location/Qualifiers
     source          1..4930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2941..2960
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3175..3763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3255..3274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3934..4725,4726..4794))
                     /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     4557..4576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4795..4899)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattccatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtggagctt
      241 actgagactc ttcggggaat tcctgtgagg gtctagtaac tgcctgagaa aatatcttgt
      301 ctgaccccgg ttatattttt ccttcggtag ggattggact ttctgaagga cgttgtgatc
      361 caaaggaagg aggccggagg tctctacttc ccatacagca ggtaactaag ttgtctgtag
      421 cagactgtct acaggcatat cgtgagacga cccaggcgtc cctggggtca gagaggacct
      481 tgcctgcaag tccgggggcg gggcctgagt cagtctcgcc agctgccggt ctttcggggg
      541 ctccgtaact ttctatccgt ccgcgtcagc gccttgccac cctcatctcc aatatggtat
      601 ggcggccctt ccatgatccc cgcctctccc agaagccctg actcctcctg ctttgcgccg
      661 tgcttttcct ctgtagctcc cttgcttccc ccagcctcgg gtgtgggtgt ctaggccggg
      721 gttctggggc aggcctgccg cgctcacccg tctgtctgct tgtctccctc tacagggatc
      781 gggtggtacc agtggcatga gcgagctgat taaggagaac atgcacatga agctgtacat
      841 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta
      901 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
      961 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg
     1021 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     1081 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acgtaagtat
     1141 gaaattcagg gatacggcca ccttgttggc cggtaaccta caaacgggtg gaggatcacc
     1201 ccacccgaca cgcaccgatg ctctccgagg aggcacaagg gtggaggaac accccaccct
     1261 ccaggcaccg atgctctccg aggaggcaaa cagaagcacc atcagggctt ctgctacgca
     1321 ccgatgctct ccgaggagga acaaggaatc accatcaggg attccctgtg gcaccgatgc
     1381 tctccgagga ggacgacggg aggacgatca cgcctcccga atagcaccga tgctctccga
     1441 ggaggtcagt gcgtggagca tcagcccacg cagccagcac cgatgctctc cgaggaggct
     1501 ttcgcgaaga gcatcagcct tcgcgccatg caccgatgct ctccgaggag gcctacagga
     1561 acagcatcag cgttcctgcc cagcaccgat gctctccgag gaggtgtccc catgcagcat
     1621 cagcgcatgg gccccgcacc gatgctctcc gaggaggcac atccgagcac caacagggct
     1681 cggagtgtgc accgatgctc tccgaggagg accctccatg gaccatcagg ccatggactc
     1741 tgcaccgatg ctctccgagg aggcttctcg aagcagcatc agcgcttcga aacagcaccg
     1801 atgctctccg aggaggttct ccaccttgtt ggcatatttg ccaaatagtg gaaatgtgaa
     1861 gtactgacaa aacttttccc tttttcaatc taatagggct gcctcatcta caacgtcaag
     1921 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     1981 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     2041 gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc
     2101 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     2161 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     2221 tactgcgacc tccctagcaa actggggcac aagcttaatg gaaccggtgc tggatccggt
     2281 cctggtccga cccccagtgg cactaacgtg ggatcctcag ggcgctctcc cagcaaagca
     2341 gtggccgccc gggcggcggg atccactgtc cggcagaggt aaggaaccct gcagttcgtt
     2401 cgcttccaga ctcggagata ggacccagaa cctcgctgat tctggggtgg agaccctagc
     2461 atgtgaagat tgacaaaggc aaaatgagct tctagtgacg tggccgtggg agtagttaaa
     2521 ggccttttgg gaggaaggcg acattttttt tctcgttgct cagtttaggg cactactctt
     2581 aaaaaaggaa agttaacaaa ctggaataga gtcagagata actttgagaa aaccgatgtc
     2641 attaaactgg tgtctctgga cctgaggttt gcactcacat ttccatctgg cggccccata
     2701 agcaatctgt cctacagata actcgtccta cacaaaactt agtctctttt cagctcagct
     2761 ctctcactct caattatatc tccttacttc catatggcac tgttgtacga tatcggagct
     2821 tactgagact cttcgggccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     2881 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     2941 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     3001 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     3061 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg acggtctcga ctcacatgtg
     3121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3601 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc
     4081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4921 acctgacgtc
//
