LOCUS       sequence-458269-        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for human MeCP2 C-terminally tagged to 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  AUTHORS   Schmidt A, Zhang H, Schmitt S, Rausch C, Popp O, Chen J, Cmarko D, 
            Butter F, Dittmar G, Lermyte F, Cardoso MC
  TITLE     The Proteomic Composition and Organization of Constitutive 
            Heterochromatin in Mouse Tissues.
  JOURNAL   Cells. 2024 Jan 11;13(2):139. doi: 10.3390/cells13020139.
  PUBMED    38247831
REFERENCE   2  (bases 1 to 6299)
  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: "Cells."; 
            date: "2024-01-11"; volume: "13(2):139. doi"; pages: " 
            10.3390/cells13020139"
COMMENT     Sequence Label: pHThMeCP2 (pc3972)
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3284..3304
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    3790..3911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3827..3846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3881..3900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3918..4373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4055..4076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4267..4286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4400..4504
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4506..4863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4506..4697
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4529..4549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4714..4849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4898..5692
                     /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             4898..5692
                     /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(4952..4971)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5562..5581
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5880..5899)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5924..5971
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgatggga tccgaaatcg gtacaggctt
     1321 ccccttcgac ccccattatg tggaagtcct gggcgagcgt atgcactacg tcgatgttgg
     1381 accgcgggat ggcacgcctg tgctgttcct gcacggtaac ccgacctcgt cctacctgtg
     1441 gcgcaacatc atcccgcatg tagcaccgag tcatcggtgc attgctccag acctgatcgg
     1501 gatgggaaaa tcggacaaac cagacctcga ttatttcttc gacgaccacg tccgctacct
     1561 cgatgccttc atcgaagcct tgggtttgga agaggtcgtc ctggtcatcc acgactgggg
     1621 ctcagctctc ggattccact gggccaagcg caatccggaa cgggtcaaag gtattgcatg
     1681 tatggaattc atccggccta tcccgacgtg ggacgaatgg ccagaattcg cccgtgagac
     1741 cttccaggcc ttccggaccg ccgacgtcgg ccgagagttg atcatcgatc agaacgcttt
     1801 catcgagggt gcgctcccga tgggggtcgt ccgtccgctt acggaggtcg agatggacca
     1861 ctatcgcgag cccttcctca agcctgttga ccgagagcca ctgtggcgat tccccaacga
     1921 gctgcccatc gccggtgagc ccgcgaacat cgtcgcgctc gtcgaggcat acatgaactg
     1981 gctgcaccag tcacctgtcc cgaagttgtt gttctggggc acacccggcg tactgatccc
     2041 cccggccgaa gccgcgagac ttgccgaaag cctccccaac tgcaagacag tggacatcgg
     2101 cccgggattg ttcttgctcc aggaagacaa cccggacctt atcggcagtg agatcgcgcg
     2161 ctggctcccc gggctggccg gctccggaat ggtagctggg atgttagggc tcagggaaga
     2221 aaagtcagaa gaccaggacc tccagggcct caaggacaaa cccctcaagt ttaaaaaggt
     2281 gaagaaagat aagaaagaag agaaagaggg caagcatgag cccgtgcagc catcagccca
     2341 ccactctgct gagcccgcag aggcaggcaa agcagagaca tcagaagggt caggctccgc
     2401 cccggctgtg ccggaagctt ctgcctcccc caaacagcgg cgctccatca tccgtgaccg
     2461 gggacccatg tatgatgacc ccaccctgcc tgaaggctgg acacggaagc ttaagcaaag
     2521 gaaatctggc cgctctgctg ggaagtatga tgtgtatttg atcaatcccc agggaaaagc
     2581 ctttcgctct aaagtggagt tgattgcgta cttcgaaaag gtaggcgaca catccctgga
     2641 ccctaatgat tttgacttca cggtaactgg gagagggagc ccctcccggc gagagcagaa
     2701 accacctaag aagcccaaat ctcccaaagc tccaggaact ggcagaggcc ggggacgccc
     2761 caaagggagc ggcaccacga gacccaaggc ggccacgtca gagggtgtgc aggtgaaaag
     2821 ggtcctggag aaaagtcctg ggaagctcct tgtcaagatg ccttttcaaa cttcgccagg
     2881 gggcaaggct gaggggggtg gggccaccac atccacccag gtcatggtga tcaaacgccc
     2941 cggcaggaag cgaaaagctg aggccgaccc tcaggccatt cccaagaaac ggggccgaaa
     3001 gccggggagt gtggtggcag ccgctgccgc cgaggccaaa aagaaagccg tgaaggagtc
     3061 ttctatccga tctgtgcagg agaccgtact ccccatcaag aagcgcaaga cccgggagac
     3121 ggtcagcatc gaggtcaagg aagtggtgaa gcccctgctg gtgtccaccc tcggtgagaa
     3181 gagcgggaaa ggactgaaga cctgtaagag ccctgggcgg aaaagcaagg agagcagccc
     3241 caaggggcgc agcagcagcg cctcctcacc ccccaagaag gagcaccacc accatcacca
     3301 ccactcagag tccccaaagg cccccgtgcc actgctccca cccctgcccc cacctccacc
     3361 tgagcccgag agctccgagg accccaccag cccccctgag ccccaggact tgagcagcag
     3421 cgtctgcaaa gaggagaaga tgcccagagg aggctcactg gagagcgacg gctgccccaa
     3481 ggagccagct aagactcagc ccgcggttgc caccgccgcc acggccgcag aaaagtacaa
     3541 acaccgaggg gagggagagc gcaaagacat tgtttcatcc tccatgccaa ggccaaacag
     3601 agaggagcct gtggacagcc ggacgcccgt gaccgagaga gttagctgac tttacacgga
     3661 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     3721 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3781 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3841 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3901 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3961 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4021 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4081 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4141 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4201 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4261 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4321 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4381 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4441 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4501 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4561 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4621 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4681 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4741 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4801 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4861 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4921 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4981 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5041 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     5101 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5161 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5221 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5281 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5341 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5401 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5461 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5521 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5581 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5641 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5701 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5761 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5821 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5881 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5941 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     6001 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     6061 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     6121 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     6181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6241 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
