LOCUS       sequence-386431-        4223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid used for building MED6-mEmerald knock-in mESCs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4223)
  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 4223)
  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-mEmerald donor
FEATURES             Location/Qualifiers
     source          1..4223
                     /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             join(2807..2809,2810..3523)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLGKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2849..2870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3110..3129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3457..3478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
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 tatgatagta 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 ggttcggtga gcaagggcga
     2821 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2881 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2941 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttgac
     3001 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3061 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3121 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3181 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3241 caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca aggtgaactt
     3301 caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3361 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3421 caagctgggc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3481 cgccgccggg atcactctcg gcatggacga gctgtacaag tgaatgttgg acgaggaaga
     3541 agctggccag ctccttgtca gtcgctatac ctcagcaccg tgggaggctc ttgagcctgg
     3601 aaacactatc gcagtttgta cggaatgcac ttgttcttta caaaaggacg ttctccctac
     3661 catgttttct atgtgaatat ttcttaaata tcttttgtaa tctttcctct actggacttg
     3721 ggcagtgcac ctaactcaca gatatgccag tatatatctt taaaacaata aagttatttg
     3781 tatatattac tcatcatgcc tttcccttgt ctctgagata tgttaatgga ataattaagc
     3841 atgtcttgta ctatttggct attcctctat atgacagtaa tatatttatt gaataattga
     3901 aaggtaaagg aaaactcact tgtttgtatc attaccgtcc tatagattct tatcatgttc
     3961 gtctgttgtg agcacttgtc agtgtttgct agtgtgtgga atgtaattct ggcgatctgt
     4021 agcttaggtt cttaactatg tcctgagtta ttttggagtt tttcccattg gtatatttta
     4081 gacaggacta tattgactgt gtttgtgtat agtgtcgtgt gcatggtcca tggtttgggt
     4141 cacagcagtc tctgcgtcct cagggctctt actctatatt gtcaaagtgt ttttctccag
     4201 tgctgctgac tccagttttt cag
//
