LOCUS       sequence-241033-        6569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-tagged vector expressing secreted isoform of Dlk1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  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 6569)
  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-S-Dlk1
FEATURES             Location/Qualifiers
     source          1..6569
                     /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             1921..1992
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    2028..2600
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2195..2212)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2422..2441
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2606..3325
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEGMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3356..3376)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3356..3374)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3394..3411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3400..3511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3620..4075)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3757..3778)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3969..3988
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4102..4206
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4208..4399
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4231..4251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4396..4530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4579..5373
                     /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             4579..5373
                     /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(4633..4652)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5243..5262
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5561..5580)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5605..5652
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5981..6569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6470..6489
                     /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 attcggtgca accctggctt tcttcccgct ggacgcccgt gcccccttcg
      901 tggtccgcaa ccagaagccc agcgcagccc ccggagcagc ccctgcaccg cctccgctcc
      961 ccggaccgcg acccaggccg ccccgagatg atcgcgaccg gagccctcct gcgcgtcctc
     1021 ttgctcctgc tggctttcgg ccacagcacc tatggggctg aatgcgaccc accctgtgac
     1081 ccccagtatg gattctgcga ggctgacaat gtctgcaggt gccatgttgg ctgggagggt
     1141 cccctctgtg acaagtgtgt aactgcccct ggctgtgtca atggagtctg caaggaacca
     1201 tggcagtgca tctgcaagga tggctgggac gggaaattct gcgaaataga cgttcgggct
     1261 tgcacctcaa ccccctgcgc caacaatgga acttgcgtgg acctggagaa aggccagtac
     1321 gaatgctcct gcacacctgg gttctctgga aaggactgcc agcacaaggc tgggccctgc
     1381 gtgatcaatg gttctccctg ccagcacgga ggcgcctgcg tggatgatga gggccaggcc
     1441 tcgcatgctt cctgcctgtg cccccctggc ttctcaggca acttctgtga gatcgtagcc
     1501 gcaaccaaca gctgtacccc taacccatgc gagaacgatg gcgtctgcac cgacatcggg
     1561 ggtgacttcc gttgccgctg cccagctgga ttcgtcgaca agacctgcag ccgcccggtg
     1621 agcaactgcg ccagtggccc gtgccagaac gggggcacct gcctccagca cacccaggtg
     1681 agcttcgagt gtctgtgcaa gcccccgttc atgggtccca cgtgcgcgaa gaagcgcggg
     1741 gctagccccg tgcaggtcac ccacctgccc agcggctatg ggctcaccta ccgcctgacc
     1801 cccggggtgc acgagctgcc tgttcagcag cccgagcaac acatcctgaa ggtgtccatg
     1861 aaagagctca acaagagtac ccctctcctc accgaggata tccatcacac tggcggccgc
     1921 gactacaagg atgacgatga caaggattac aaagacgacg atgataagga ctataaggat
     1981 gatgacgaca aataatagca attcctcgac gactgcatag ggttaccccc ctctccctcc
     2041 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2101 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2161 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2221 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2281 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2341 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2401 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2461 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     2521 tgtgtttagt cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     2581 tttgaaaaac acgatgataa tgaccatggt gagcaagcag atcctgaaga acaccggcct
     2641 gcaggagatc atgagcttca aggtgaacct ggagggcgtg gtgaacaacc acgtgttcac
     2701 catggagggc tgcggcaagg gcaacatcct gttcggcaac cagctggtgc agatccgcgt
     2761 gaccaagggc gcccccctgc ccttcgcctt cgacatcctg agccccgcct tccagtacgg
     2821 caaccgcacc ttcaccaagt accccgagga catcagcgac ttcttcatcc agagcttccc
     2881 cgccggcttc gtgtacgagc gcaccctgcg ctacgaggac ggcggcctgg tggagatccg
     2941 cagcgacatc aacctgatcg aggggatgtt cgtgtaccgc gtggagtaca agggccgcaa
     3001 cttccccaac gacggccccg tgatgaagaa gaccatcacc ggcctgcagc ccagcttcga
     3061 ggtggtgtac atgaacgacg gcgtgctggt gggccaggtg atcctggtgt accgcctgaa
     3121 cagcggcaag ttctacagct gccacatgcg caccctgatg aagagcaagg gcgtggtgaa
     3181 ggacttcccc gagtaccact tcatccagca ccgcctggag aagacctacg tggaggacgg
     3241 cggcttcgta gagcagcacg agaccgccat cgcccagctg accagcctgg gcaagcccct
     3301 gggcagcctg cacgagtggg tgtaagctcg agcatgcatc tagagggccc tattcccttt
     3361 agtgagggtt aattgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3421 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actctcactg
     3481 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3541 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3601 ctggggcgcc ggcgcgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3661 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3721 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3781 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3841 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3901 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3961 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4021 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4201 aagaatcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac catagtcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4441 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4501 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4561 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4621 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4681 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4741 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4801 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4861 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4921 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4981 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5041 caggatgatc tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5101 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5161 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5221 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc
     5281 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5341 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5401 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5461 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5521 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5581 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5641 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5701 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5761 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5821 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5881 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6121 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6541 gctcgtcagg ggggcggagc ctatggaaa
//
