LOCUS       sequence-458344-        5143 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid of MBD2-C∆CC. C-terminus of MBD2 
            lacking the coiled coil domain: aa 363-396. C-terminal tagged to 
            GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5143)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pmMBD2-C∆CC-G pc5091
FEATURES             Location/Qualifiers
     source          1..5143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    463..584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(474..493)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     528..547
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(695..1408,1409..1411,1412..1414))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(698..1408,1409..1411,1412..1414))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(740..761)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1089..1108
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1348..1369
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1947..2150)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1976..1996)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2151..2454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2629..3217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2709..2728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3546..3593
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3618..3637
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3825..4619)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3825..4619)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3936..3955)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4546..4565
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4654..5011)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4668..4803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4820..5011
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4968..4988
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5013..5117)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttacgc cttaagatac attgatgagt
      481 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
      541 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
      601 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
      661 tctacaaatg tggtatggct gattatgatc tagattactt gtacagctcg tccatgccga
      721 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
      781 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
      841 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
      901 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat
      961 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     1021 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     1081 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     1141 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     1201 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     1261 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     1321 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     1381 ccccggtgaa cagctcctcg cccttgctca ccatggcgcg gccgcacgcc tcatctccat
     1441 cgtccatgtc aatgtctact tcctccgtgt ccgcagcccg gcagaggggt tgagatgtgt
     1501 taagccaaac agcagggttc ttttccacgg cagcagagac ttgtcctgtg atgggcgcag
     1561 agcttgtgtg taaagcactg gccacagcag acagaagggt ctcgtcattg ctacctggac
     1621 cgactccttg aagaccttta ggtagctcca tggtttttat aatttgttct gttacatctg
     1681 atgcgctaag tccttgtagc ctcttctccc agaaaagctg acgtggttgt tcattcatcc
     1741 gctgggggtc tgacttcacc ttattgctcg ggtggttcgt gaatttggtt actggttgct
     1801 tgaaaattga tgcagtttgt ctaattggca atgttgtgtt caggtctggt ttacccttgt
     1861 tcatgtcgac gtaggccttt gaattcgaag cttgagctcg agatctgagt ccggtagcgc
     1921 tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc
     1981 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     2041 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     2101 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     2161 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     2221 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     2281 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     2341 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     2401 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     2461 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actaatgcat
     2521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3361 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     3421 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     3481 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     3541 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     3601 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     3661 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     3721 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     3781 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     3841 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     3901 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     3961 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     4021 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     4081 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     4141 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     4201 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     4261 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     4321 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     4381 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     4441 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4501 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4561 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     4621 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     4681 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     4741 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     4801 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     4861 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     4921 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     4981 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
     5041 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5101 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
