LOCUS       sequence-386442-        4122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid used for building mEmerald-MED1 knock-in mESCs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4122)
  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 4122)
  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-mEmerald-MED1 donor
FEATURES             Location/Qualifiers
     source          1..4122
                     /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             join(2737..2739,2740..3450)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2779..2800)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3040..3059)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3387..3408
                     /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 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 cagagcgtga gcaagggcga ggagctgttc
     2761 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2821 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2881 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttgac ctacggcgtg
     2941 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3001 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3061 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3121 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3181 aaggtctata tcaccgccga caagcagaag aacggcatca aggtgaactt caagacccgc
     3241 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3301 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     3361 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3421 atcactctcg gcatggacga gctgtacaag tccggtggcg gcggatctgg cggtggtggg
     3481 tccggaggag gcggttccaa ggctcagggg gaaaccgagg gtgagtgact ggggggtggc
     3541 ggggaaggag gggcggtaca ccgctgcggc gagccaatcc gagagaggag gtggggaccc
     3601 acgcccctcc agggccaccg tgcgtgggac cggatcctgg tggaatgtgc atcttagatc
     3661 ccgaccgagc cctacagatg ctggagcgcc tcctccgggg cgctgctccc ccgtctcctc
     3721 cactcgtgaa aggttttctg cgggacgagg gacgaggaag ttgagcctca gggtactttg
     3781 agcttagccc ggtgggaccc aaggagaggg aaactgaaac ttgttgatgc tttgttccag
     3841 gagagatcgc tgaagatgac actagaacaa tgtaatgtca aagtaatgga gggtgtttta
     3901 gccggagtga cttgcttggt tcgatcttca tcatagtaaa agcgggccgt ttattacttg
     3961 gaaggtcata gtgttcaaat actgtgctgt gtctttacga actgctttca gttgtgcacg
     4021 accggtggct ttaaagcttg ggggagaggt gttcagaact ataaactggc aggaacaatc
     4081 cccaaaccag tttcccagtt gagggaagaa aagggaaggg at
//
