LOCUS       sequence-241029-        6602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-tagged vector expressing membrane-bound Dlk1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6602)
  AUTHORS   Ferron SR, Charalambous M, Radford E, McEwen K, Wildner H, Hind E, 
            Morante-Redolat JM, Laborda J, Guillemot F, Bauer SR, Farinas I, 
            Ferguson-Smith AC
  TITLE     Postnatal loss of Dlk1 imprinting in stem cells and niche astrocytes
            regulates neurogenesis.
  JOURNAL   Nature. 2011 Jul 20;475(7356):381-5. doi: 10.1038/nature10229.
  PUBMED    21776083
REFERENCE   2  (bases 1 to 6602)
  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"; journalName: 
            "Nature"; date: "2011-07-20- 20"; volume: "475"; issue: "7356"; 
            pages: "381-5"
COMMENT     Sequence Label: pIRESh-GFPII-MB-Dlk1
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        145..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..728
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..792
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        773..791
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1954..2025
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    2061..2633
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2228..2245)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2455..2474
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2639..3358
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEGMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3389..3409)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3389..3407)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3427..3444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3433..3544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3653..4108)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3811)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4002..4021
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4135..4239
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4241..4432
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4264..4284)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4429..4563
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4612..5406
                     /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             4612..5406
                     /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(4666..4685)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5276..5295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5594..5613)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5638..5685
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccag gcgcgccgcg
      121 atgtacgggc cagatttacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      661 acaactccgc cccattgaag caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      721 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
      781 ctcactatag ggagacccaa gcttctggag gcccgggctt tcagggtacc gaagaaggat
      841 ccaaggagga attcggcacg aggcgccccg gtgcaaccct ggctttcttc ccgctggacg
      901 cccgtgcccc cttcgtggtc cgcaaccaga agcccagcgc agcccccgga gcagcccctg
      961 caccgcctcc gctccccgga ccgcgaccca ggccgccccg agatgatcgc gaccggagcc
     1021 ctcctgcgcg tcctcttgct cctgctggct ttcggccaca gcacctatgg ggctgaatgc
     1081 gacccaccct gtgaccccca gtatggattc tgcgaggctg acaatgtctg caggtgccat
     1141 gttggctggg agggtcccct ctgtgacaag tgtgtaactg cccctggctg tgtcaatgga
     1201 gtctgcaagg aaccatggca gtgcatctgc aaggatggct gggacgggaa attctgcgaa
     1261 atagacgttc gggcttgcac ctcaaccccc tgcgccaaca atggaacttg cgtggacctg
     1321 gagaaaggcc agtacgaatg ctcctgcaca cctgggttct ctggaaagga ctgccagcac
     1381 aaggctgggc cctgcgtgat caatggttct ccctgccagc acggaggcgc ctgcgtggat
     1441 gatgagggcc aggcctcgca tgcttcctgc ctgtgccccc ctggcttctc aggcaacttc
     1501 tgtgagatcg tagccgcaac caacagctgt acccctaacc catgcgagaa cgatggcgtc
     1561 tgcaccgaca tcgggggtga cttccgttgc cgctgcccag ctggattcgt cgacaagacc
     1621 tgcagccgcc cggtgagcaa ctgcgccagt ggcccgtgcc agaacggggg cacctgcctc
     1681 cagcacaccc aggccatctg cttcaccatc ctgggcgtgc tcaccagcct ggtggtgctg
     1741 ggcaccgtgg ccatcgtctt tctcaacaag tgcgaaacct gggtgtccaa cctgcgctac
     1801 aaccacatgc ttcgcaagaa gaagaacctc ctgttgcagt ataacagcgg cgaggagctg
     1861 gcggtcaata tcatcttccc cgagaagatt gacatgacca ctttcaacaa ggaggctggt
     1921 gatgaggaga tcatccatca cactggcggc cgcgactaca aggatgacga tgacaaggat
     1981 tacaaagacg acgatgataa ggactataag gatgatgacg acaaataata gcaattcctc
     2041 gacgactgca tagggttacc cccctctccc tccccccccc ctaacgttac tggccgaagc
     2101 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     2161 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     2221 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     2281 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     2341 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     2401 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     2461 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     2521 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc
     2581 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatgaccat
     2641 ggtgagcaag cagatcctga agaacaccgg cctgcaggag atcatgagct tcaaggtgaa
     2701 cctggagggc gtggtgaaca accacgtgtt caccatggag ggctgcggca agggcaacat
     2761 cctgttcggc aaccagctgg tgcagatccg cgtgaccaag ggcgcccccc tgcccttcgc
     2821 cttcgacatc ctgagccccg ccttccagta cggcaaccgc accttcacca agtaccccga
     2881 ggacatcagc gacttcttca tccagagctt ccccgccggc ttcgtgtacg agcgcaccct
     2941 gcgctacgag gacggcggcc tggtggagat ccgcagcgac atcaacctga tcgaggggat
     3001 gttcgtgtac cgcgtggagt acaagggccg caacttcccc aacgacggcc ccgtgatgaa
     3061 gaagaccatc accggcctgc agcccagctt cgaggtggtg tacatgaacg acggcgtgct
     3121 ggtgggccag gtgatcctgg tgtaccgcct gaacagcggc aagttctaca gctgccacat
     3181 gcgcaccctg atgaagagca agggcgtggt gaaggacttc cccgagtacc acttcatcca
     3241 gcaccgcctg gagaagacct acgtggagga cggcggcttc gtagagcagc acgagaccgc
     3301 catcgcccag ctgaccagcc tgggcaagcc cctgggcagc ctgcacgagt gggtgtaagc
     3361 tcgagcatgc atctagaggg ccctattccc tttagtgagg gttaattgct agagctcgct
     3421 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3481 cttccttgac cctggaaggt gccactctca ctgtcctttc ctaataaaat gaggaaattg
     3541 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3601 agggggagga ttgggaagac aatagcaggc atgctggggc gccggcgcgc gtaaattgta
     3661 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3721 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3781 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3841 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3901 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3961 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4021 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4081 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4141 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4201 ccctgataaa tgcttcaata atattgaaaa aggaagaatc ctgaggcgga aagaaccagc
     4261 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4321 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4441 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4501 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4561 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4621 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4681 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4741 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4801 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4861 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4921 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4981 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5041 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agaacatcag
     5101 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5161 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5221 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5281 gctacccgtg atattgctga agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5341 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5401 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5461 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5521 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5581 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5641 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5701 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5761 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5821 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aa
//
