LOCUS       sequence-386849-        6472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The miRFP670-gene is  ligated  c-terminally to the rat meCP2-gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6472)
  AUTHORS   Chagin VO, Reinhart B, Becker A, Mortusewicz O, Jost KL, Rapp A, 
            Leonhardt H, Cardoso MC
  TITLE     Processive DNA synthesis is associated with localized decompaction 
            of constitutive heterochromatin at the sites of DNA replication and 
            repair.
  JOURNAL   Nucleus. 2019 Dec;10(1):231-253. doi: 10.1080/19491034.2019.1688932.
  PUBMED    31744372
REFERENCE   2  (bases 1 to 6472)
  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"; doi: 
            "10.1080/19491034.2019"; journalName: "Nucleus"; date: "2019-12"; 
            volume: "10"; issue: "1"; pages: "231-253"
COMMENT     Sequence Label: pMeCP2miRFP670 pc3942
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        417..620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     571..591
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1316..2173
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     regulatory      2237..2246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2243..3190
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    3312..3433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3349..3368)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3403..3422
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3440..3895)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3577..3598)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3789..3808
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3922..4026
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4028..4385
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4028..4219
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4051..4071)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4236..4371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4420..5214
                     /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             4420..5214
                     /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(4474..4493)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5084..5103
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5402..5421)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5446..5493
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5822..6410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6311..6330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
       61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      601 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      661 agatctcgag ctccaccgcg gtggcggccg ctctagcatg gtagctggga tgttagggct
      721 caggaaggaa aagtcagaag accaggatct ccagggcctc aaagagaaac ccctgaagtt
      781 taagaaggtg aagaaagaca agaaggaaga caaagagggc aaacatgaac cactacagcc
      841 ttcagcccac cattctgcag agccagcaga ggcaggcaaa gcagagacat cagaaagctc
      901 aggctctgcc ccagcagtac cagaagcctc tgcttctccc aaacagcgac gttccatcat
      961 tcgtgaccgg ggacctatgt atgatgaccc caccttgcct gaaggttgga cgcgaaagct
     1021 taaacagagg aagtctggtc gctctgctgg aaagtatgat gtatatttga tcaatcccca
     1081 gggaaaagcc tttcgctcta aagtagaatt gattgcatat tttgaaaagg tgggagacac
     1141 ctccttggac cctaatgatt ttgacttcac tgtaactggg agagggagcc cttccaggag
     1201 agaacagaaa ccacctaaga agcccaaatc tcccaaagct ccaggaactg gcaggggtcg
     1261 gggacgcccg aaagggagcg gcactgggag accaaaggca gcagcatcag aaggtgttca
     1321 agtgaaaagg gtcctggaga agagccctgg gaaacttctc gtcaagatgc ctttccaagc
     1381 atcacctggg ggtaagggtg agggaggtgg ggctaccaca tctgcgcagg tcatggtgat
     1441 caaacgccct ggcagaaagc gaaaagctga agctgacccc caggccattc ctaagaaacg
     1501 gggtagaaag cctgggagtg tggtggcagc tgctgctgca gaggccaaaa agaaagctgt
     1561 gaaggaatct tctatacggt ctgtgcagga gactgtgctc cccatcaaga agcgcaagac
     1621 ccgggaaacc gtcagcattg aggtcaagga ggtggtgaag cccctgctgg tgtctacact
     1681 tggtgagaag agtggaaagg gactgaagac atgcaagagc cctgggcgta aaagcaagga
     1741 gagcagcccc aaggggcgca gcagcagtgc ctcctcacca cctaagaagg agcaccatca
     1801 tcaccaccat cacgcagagt ccccaaaggc ccccatgcca ttgcttccac ctccaccccc
     1861 acctgagcct cagagctctg aggaccccat cagcccccct gagcctcagg acttgagcag
     1921 cagcatctgc aaagaggaga agatgccccg agcaggctca ctggaaagcg atggctgccc
     1981 caaggagcca gctaagactc agcccatggt tgctgccgcc gccaccacca ccaccaccac
     2041 caccaccaca gttgcagaaa agtacaaaca ccgaggggag ggagagcgca aagacattgt
     2101 ttcatcctcc atgccgaggc caaacagaga ggagcctgtg gacagccgga cgcccgtgac
     2161 cgagagagtt agcaggaatt cgatatcaag cttatcgata ccgtcgacgg taccgcgggc
     2221 ccgggatcca ccggtcgcca ccatggtagc aggtcatgcc tctggcagcc ccgcattcgg
     2281 gaccgcctct cattcgaatt gcgaacatga agagatccac ctcgccggct cgatccagcc
     2341 gcatggcgcg cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa
     2401 cgccgcggaa tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg
     2461 cgatctgttg atcaagatcc tgccgcatct cgatcccacc gccgaaggca tgccggtcgc
     2521 ggtgcgctgc cggatcggca atccctctac ggagtactgc ggtctgatgc atcggcctcc
     2581 ggaaggcggg ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac
     2641 gctggcgccg gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga
     2701 caccgtgctg ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga
     2761 tgagcaaggc cacggcctgg tattctccga gtgccatgtg cctgggctcg aatcctattt
     2821 cggcaaccgc tatccgtcgt cgactgtccc gcagatggcg cggcagctgt acgtgcggca
     2881 gcgcgtccgc gtgctggtcg acgtcaccta tcagccggtg ccgctggagc cgcggctgtc
     2941 gccgctgacc gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgtg
     3001 ccatctgcag ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt
     3061 cggcggcaag ctgtggggcc tggttgtctg tcaccattat ctgccgcgct tcatccgttt
     3121 cgagctgcgg gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct
     3181 tgagagctaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     3241 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3301 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3361 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3421 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3481 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3541 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3601 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3661 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3721 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3781 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3841 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3901 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3961 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4021 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4141 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4201 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4321 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4381 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4441 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4501 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4561 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4621 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4681 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4741 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4801 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4861 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4921 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4981 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5041 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5101 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5161 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5221 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5281 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5341 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5401 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5461 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5521 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5581 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5641 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5701 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5761 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5821 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5881 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5941 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6001 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6061 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6121 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6181 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6241 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6301 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6361 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6421 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
