LOCUS       sequence-438164-        7680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP in Newborn neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7680)
  AUTHORS   Luikart BW, Bensen AL, Washburn EK, Perederiy JV, Su KG, Li Y, 
            Kernie SG, Parada LF, Westbrook GL
  TITLE     miR-132 mediates the integration of newborn neurons into the adult 
            dentate gyrus.
  JOURNAL   PLoS One. 2011;6(5):e19077. doi: 10.1371/journal.pone.0019077. Epub 
            2011 May 17.
  PUBMED    21611182
REFERENCE   2  (bases 1 to 7680)
  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: "PLoS One.";
            date: "2011"; volume: "6(5):e19077. doi"; pages: " 
            10.1371/journal.pone.0019077. Epub 2011 May 17"
COMMENT     Sequence Label: pRubi-G
FEATURES             Location/Qualifiers
     source          1..7680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             586..761
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    824..1023
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1224..1640
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1570..1592
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1608..1624
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1711..2922
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2759..2778
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      2970..2979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2976..2978,2979..2981,2982..3695)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2976..2978,2979..2981,2982..3692)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3021..3042)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3282..3301)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3629..3650
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3736..4324
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3789..3809)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4207..4218)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4327..4343)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4328..4344)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4459..4884
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4987..5009)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5145..5165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5150..5479
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5330..5465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5501..5520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5601..5618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5772..6360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5852..5871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6531..7322,7323..7391))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7154..7173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7392..7496)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7564..7582
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata aaagagccca
      601 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
      661 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
      721 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
      781 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
      841 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
      901 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
      961 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     1021 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     1081 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     1141 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     1201 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     1261 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     1321 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     1381 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     1441 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     1501 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     1561 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     1621 tcactccttc tctaggcgcc ggaattgaag atctcacaca gcggccgcac acacttcgaa
     1681 ccttaattaa cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg gcgcctcccg
     1741 cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc
     1801 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
     1861 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
     1921 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
     1981 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
     2041 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
     2101 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
     2161 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
     2221 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     2281 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     2341 ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg
     2401 agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     2461 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     2521 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     2581 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     2641 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     2701 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     2761 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     2821 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa
     2881 ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagcttg ggctgcaggt
     2941 cgactctaga ggatccccgg gtaccggtcg ccaccatggt gagcaagggc gaggagctgt
     3001 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3061 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     3121 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     3181 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3241 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3301 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3361 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3421 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3481 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3541 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     3601 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3661 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agaattcgat
     3721 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3781 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3841 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3901 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3961 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4021 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4081 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     4141 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4201 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4261 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4321 ccgcatcgat accgtcgacc tcgagatatc agtggtccag gctctagttt tgactcaaca
     4381 atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag
     4441 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     4501 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     4561 gtcaggaaca gatggaacag ggtcgaccct agagaaccat cagatgtttc cagggtgccc
     4621 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     4681 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     4741 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     4801 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     4861 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
     4921 ggaccaccga cccaccaccg ggaggtaagc tggctgcctc gcgcgtttcg gtgatgacgg
     4981 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5041 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     5101 catgacccag tcacgtagcg atagcggagt gtagatccgg ctgtggaatg tgtgtcagtt
     5161 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5221 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5281 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5341 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5401 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5461 aggcctaggc ttttgcaaaa agcttactgg cttaactatg cggcatcaga gcagattgta
     5521 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5581 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5641 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6181 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6841 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7141 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7381 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7441 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7501 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7561 aaaaataggc gtatcacgag gccctttcgt cttcaagaat tagcttggcc attgcatacg
     7621 ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt accgccatgt
//
