LOCUS       sequence-386427-        4394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid used for building MED6-Halo knock-in mESCs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4394)
  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 4394)
  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-MED6-Halo donor
FEATURES             Location/Qualifiers
     source          1..4394
                     /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             2807..3691
                     /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 ttaagccctg tcccaggtat ggtttatttg ctgctcttat taaacttaaa
     2101 gtaagatcag gcagctgtgt aatgttctaa cagcagcggc atgggagatg agctgcagcc
     2161 agtttgtccc ttaggcatct cagtatagtt tgccattcct aacctttgac ttgttgaatg
     2221 caagcagata gtggctctgt ggaagtgacc aaagcatttg gaggtagagt ctgggtggaa
     2281 agattgggaa gagcaaggag gccaaagtga catgatagta ggtttcacca ttgtgaacag
     2341 gtgttgagct gtctgtgatg ttgccagttt ttgtggcttg ttcacacata atgtctcatt
     2401 ttagtacaaa catgtttgcc tagcattata aaattttttg atttattttt tttaggttta
     2461 taagagaatt ttaaaatatg tttacctggg ggaaacaact aaatattcaa ccacataagc
     2521 aattaaaaat gaatttgaaa gagtaaacct ttaatgttaa ccagtgttta aaattatgtc
     2581 tttcttctac cttcatttgt cttgacaaaa ttcctgcttt tgctgtctag tggatcaggc
     2641 aaaaaaagag gcagagcctt tgccagaaac tgtaaaatct gaggagaagg agagcacaaa
     2701 gaacattcag cagactgtga gcactaaagg tccccctgag aaacgaatga gactccagtc
     2761 cggtggcggc ggatctggcg gtggtgggtc cggaggaggc ggttcggaaa tcggtactgg
     2821 ctttccattc gacccccatt atgtggaagt cctgggcgag cgcatgcact acgtcgatgt
     2881 tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt aacccgacct cctcctacgt
     2941 gtggcgcaac atcatcccgc atgttgcacc gacccatcgc tgcattgctc cagacctgat
     3001 cggtatgggc aaatccgaca aaccagacct gggttatttc ttcgacgacc acgtccgctt
     3061 catggatgcc ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca ttcacgactg
     3121 gggctccgct ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca aaggtattgc
     3181 atttatggag ttcatccgcc ctatcccgac ctgggacgaa tggccagaat ttgcccgcga
     3241 gaccttccag gccttccgca ccaccgacgt cggccgcaaa ctgatcatcg atcagaacgt
     3301 ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga
     3361 ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa
     3421 cgagctgcca atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga
     3481 ctggctgcac cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat
     3541 cccaccggcc gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat
     3601 cggcccgggt ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc
     3661 gcgctggctg tctactctgg agatttccgg ttgaatgttg gacgaggaag aagctggcca
     3721 gctccttgtc agtcgctata cctcagcacc gtgggaggct cttgagcctg gaaacactat
     3781 cgcagtttgt acggaatgca cttgttcttt acaaaaggac gttctcccta ccatgttttc
     3841 tatgtgaata tttcttaaat atcttttgta atctttcctc tactggactt gggcagtgca
     3901 cctaactcac agatatgcca gtatatatct ttaaaacaat aaagttattt gtatatatta
     3961 ctcatcatgc ctttcccttg tctctgagat atgttaatgg aataattaag catgtcttgt
     4021 actatttggc tattcctcta tatgacagta atatatttat tgaataattg aaaggtaaag
     4081 gaaaactcac ttgtttgtat cattaccgtc ctatagattc ttatcatgtt cgtctgttgt
     4141 gagcacttgt cagtgtttgc tagtgtgtgg aatgtaattc tggcgatctg tagcttaggt
     4201 tcttaactat gtcctgagtt attttggagt ttttcccatt ggtatatttt agacaggact
     4261 atattgactg tgtttgtgta tagtgtcgtg tgcatggtcc atggtttggg tcacagcagt
     4321 ctctgcgtcc tcagggctct tactctatat tgtcaaagtg tttttctcca gtgctgctga
     4381 ctccagtttt tcag
//
