LOCUS       sequence-243672-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-tagged vector expressing full length isoform of Dlk1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  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 6827)
  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-SM-Dlk1
FEATURES             Location/Qualifiers
     source          1..6827
                     /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             2179..2250
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    2286..2858
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2453..2470)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2680..2699
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2864..3583
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEGMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3614..3634)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3614..3632)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3652..3669)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3658..3769
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3878..4333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4015..4036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4227..4246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4360..4464
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4466..4657
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4489..4509)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4654..4788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4837..5631
                     /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             4837..5631
                     /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(4891..4910)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5501..5520
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5819..5838)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5863..5910
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6239..6827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6728..6747
                     /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 aggtgagctt cgagtgtctg tgcaagcccc cgttcatggg tcccacgtgc
     1741 gcgaagaagc gcggggctag ccccgtgcag gtcacccacc tgcccagcgg ctatgggctc
     1801 acctaccgcc tgacccccgg ggtgcacgag ctgcctgttc agcagcccga gcaacacatc
     1861 ctgaaggtgt ccatgaaaga gctgaacaag agtacccctc tcctcaccga gggacaggcc
     1921 atctgcttca ccatcctggg cgtgctcacc agcctggtgg tgctgggcac cgtggccatc
     1981 gtctttctca acaagtgcga aacctgggtg tccaacctgc gctacaacca catgcttcgc
     2041 aagaagaaga acctcctgtt gcagtataac agcggcgagg agctggcggt caatatcatc
     2101 ttccccgaga agattgacat gaccactttc aacaaggagg ctggtgatga ggagatcatc
     2161 catcacactg gcggccgcga ctacaaggat gacgatgaca aggattacaa agacgacgat
     2221 gataaggact ataaggatga tgacgacaaa taatagcaat tcctcgacga ctgcataggg
     2281 ttacccccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     2341 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     2401 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     2461 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     2521 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     2581 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     2641 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2701 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2761 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc
     2821 acggggacgt ggttttcctt tgaaaaacac gatgataatg accatggtga gcaagcagat
     2881 cctgaagaac accggcctgc aggagatcat gagcttcaag gtgaacctgg agggcgtggt
     2941 gaacaaccac gtgttcacca tggagggctg cggcaagggc aacatcctgt tcggcaacca
     3001 gctggtgcag atccgcgtga ccaagggcgc ccccctgccc ttcgccttcg acatcctgag
     3061 ccccgccttc cagtacggca accgcacctt caccaagtac cccgaggaca tcagcgactt
     3121 cttcatccag agcttccccg ccggcttcgt gtacgagcgc accctgcgct acgaggacgg
     3181 cggcctggtg gagatccgca gcgacatcaa cctgatcgag gggatgttcg tgtaccgcgt
     3241 ggagtacaag ggccgcaact tccccaacga cggccccgtg atgaagaaga ccatcaccgg
     3301 cctgcagccc agcttcgagg tggtgtacat gaacgacggc gtgctggtgg gccaggtgat
     3361 cctggtgtac cgcctgaaca gcggcaagtt ctacagctgc cacatgcgca ccctgatgaa
     3421 gagcaagggc gtggtgaagg acttccccga gtaccacttc atccagcacc gcctggagaa
     3481 gacctacgtg gaggacggcg gcttcgtaga gcagcacgag accgccatcg cccagctgac
     3541 cagcctgggc aagcccctgg gcagcctgca cgagtgggtg taagctcgag catgcatcta
     3601 gagggcccta ttccctttag tgagggttaa ttgctagagc tcgctgatca gcctcgactg
     3661 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3721 aaggtgccac tctcactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3781 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3841 aagacaatag caggcatgct ggggcgccgg cgcgcgtaaa ttgtaagcgt taatattttg
     3901 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3961 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4021 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4081 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4141 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4201 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4261 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4321 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4381 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4441 caataatatt gaaaaaggaa gaatcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4501 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4561 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4621 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccagttccg
     4681 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4741 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4801 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4861 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4921 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4981 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     5041 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5101 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5161 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5221 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5281 gaagccggtc ttgtcgatca ggatgatctg gacgaagaac atcaggggct cgcgccagcc
     5341 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5401 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5461 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5521 gctgaagaac ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5581 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5641 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5701 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5761 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5821 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5881 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5941 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     6001 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     6061 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     6121 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6181 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaa
//
