LOCUS       sequence-386426-        4308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid used for building Halo-MED1 knock-in mESCs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4308)
  AUTHORS   Dong P, Zhang S, Xie L, Wang L, Lemire AL, Lander AD, Chang HY, Liu 
            ZJ
  TITLE     Spatial organization of the 3D genome encodes gene co-expression 
            programs in single cells
  JOURNAL   bioRxiv 2022.10.26.513917
REFERENCE   2  (bases 1 to 4308)
  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: "bioRxiv 
            2022.10.26.513917"
COMMENT     Sequence Label: pminiT-Halo-MED1 donor
FEATURES             Location/Qualifiers
     source          1..4308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(116..184,185..976)
                     /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(334..353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1147..1735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1636..1655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1889..1906
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             2716..2736
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2752..3636
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
ORIGIN
        1 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
       61 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      301 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      421 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
      661 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      721 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      961 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1741 cagcaatgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1921 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gccggttcag
     1981 acaggataaa gaggaacgca gaatgttaga caacacccgc ttacgcatag ctattcagaa
     2041 atcaggccgt ttaagcaccg gttttcacta tccccttcct cattgggatt ataggagtgt
     2101 gccacttttc caggtgctca aagagcaaac attttgtttc gtttgagaca gggtttctct
     2161 gtgtagccat ggctgtcctg gaactcacac tgtagaccag gctggcctcg aactcagagt
     2221 tccgcctgtc tgctgggatc aaaggcttga gccgccacca ccgcgggacg gaagagctaa
     2281 cgcttttaat ttacaaattg tgtaactgtc gggatgcaaa cataaacttt cagagagatg
     2341 tcaactctgg gcacacaccc ttagacgcca aggtgtgaat tctgagccaa tcggagccgg
     2401 gcctgctaga gatcctgcgg ggaggacgaa atcagccaac ctccgcgaga ctttggtgcg
     2461 gttccttctt cctccgtctc acaggaacac gattaatctc gcgatctttg ggaagttccg
     2521 ttggggaaga tggcggcggc cgcgagcacc cttctcttct tgccgccggg gacttcagac
     2581 tgagccttcc ccggaagagt cggggccgct ggtgcgccgc ttccccggag cccaggctaa
     2641 cttggttccc ctctgagtgg cggggaaagg ctccccggct cgtggcggac ccaagacctc
     2701 cccgctgtca ggatggagga tctgtacttt cagagcgata acgatggatc cgaaatcggt
     2761 actggctttc cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc
     2821 gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc
     2881 tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac
     2941 ctgatcggta tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc
     3001 cgcttcatgg atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac
     3061 gactggggct ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt
     3121 attgcattta tggagttcat ccgccctatc ccgacctggg acgaatggcc agaatttgcc
     3181 cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc gcaaactgat catcgatcag
     3241 aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag
     3301 atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc
     3361 ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac
     3421 atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt
     3481 ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg
     3541 gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag
     3601 atcgcgcgct ggctgtctac tctggagatt tccggttccg gtggcggcgg atctggcggt
     3661 ggtgggtccg gaggaggcgg ttccaaggct cagggggaaa ccgagggtga gtgactgggg
     3721 ggtggcgggg aaggaggggc ggtacaccgc tgcggcgagc caatccgaga gaggaggtgg
     3781 ggacccacgc ccctccaggg ccaccgtgcg tgggaccgga tcctggtgga atgtgcatct
     3841 tagatcccga ccgagcccta cagatgctgg agcgcctcct ccggggcgct gctcccccgt
     3901 ctcctccact cgtgaaaggt tttctgcggg acgagggacg aggaagttga gcctcagggt
     3961 actttgagct tagcccggtg ggacccaagg agagggaaac tgaaacttgt tgatgctttg
     4021 ttccaggaga gatcgctgaa gatgacacta gaacaatgta atgtcaaagt aatggagggt
     4081 gttttagccg gagtgacttg cttggttcga tcttcatcat agtaaaagcg ggccgtttat
     4141 tacttggaag gtcatagtgt tcaaatactg tgctgtgtct ttacgaactg ctttcagttg
     4201 tgcacgaccg gtggctttaa agcttggggg agaggtgttc agaactataa actggcagga
     4261 acaatcccca aaccagtttc ccagttgagg gaagaaaagg gaagggat
//
