LOCUS       sequence-365453-        8194 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gli activity reporter, mouse transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8194)
  AUTHORS   Balaskas N, Ribeiro A, Panovska J, Dessaud E, Sasai N, Page KM, 
            Briscoe J, Ribes V
  TITLE     Gene regulatory logic for reading the Sonic Hedgehog signaling 
            gradient in the vertebrate neural tube.
  JOURNAL   Cell. 2012 Jan 20;148(1-2):273-84. doi: 10.1016/j.cell.2011.10.047.
  PUBMED    22265416
REFERENCE   2  (bases 1 to 8194)
  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: "Cell."; 
            date: "2012-01-20"; volume: "148(1-2):273-84. doi"; pages: " 
            10.1016/j.cell.2011.10.047"
COMMENT     Sequence Label: GBSGFP insulator
FEATURES             Location/Qualifiers
     source          1..8194
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1856..1858,1859..1861,1862..2575)
                     /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(1856..1858,1859..1861,1862..2572)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1901..1922)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2162..2181)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2509..2530
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2708..2842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2745..2764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2799..2818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(5247..5265)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5248..5265)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5273..5290)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5273..5289)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5282..5304)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5498..5517)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5617..5639
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5677..5695)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5763..5867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5868..5936,5937..6728)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6086..6105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6899..7487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7388..7407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7641..7658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7775..7796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7811..7816,7817..7834,7835..7841)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7849..7865
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7854..7876
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7873..7889
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7873..7889
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7908..7927
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7908..7926
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagcaagtta aaacccagca tggagaggag cacaaaaagg ccacagacac tgctggtccc
       61 tgtgtctgag cctgcatgtt tgatggtgtc tggatgcaag cagaaggggt ggaagagctt
      121 gcctggagag atacagctgg gtcagtagga ctgggacagg cagctggaga attgccatgt
      181 agatgttcat acaatcgtca aatcatgaag gctggaaaag ccctccaaga tccccaagac
      241 caaccccaac ccacccaccg tgcccactgg ccatgtccct cagtgccaca tccccacagt
      301 tcttcatcac ctccagggac ggtgaccccc ccacctccgt gggcagctgt gccactgcag
      361 caccgctctt tggagaaggt aaatcttgct aaatccagcc cgaccctccc ctggcacaac
      421 gtaaggccat tatctctcat ccaactccag gacggagtca gtgaggatgg ggctctagag
      481 gatccttatg acggaggcta acaagcaggg aacacccaag tagaagctgg ctgtcagatc
      541 cttatgacgg aggctaacaa gcagggaaca cccaagtaga agctggctgt cagatcctta
      601 tgacggaggc taacaagcag ggaacaccca agtagaagct ggctgtcaga tccttatgac
      661 ggaggctaac aagcagggaa cacccaagta gaagctggct gtcagatcct tatgacggag
      721 gctaacaagc agggaacacc caagtagaag ctggctgtca gatccttatg acggaggcta
      781 acaagcaggg aacacccaag tagaagctgg ctgtcagatc cttatgacgg aggctaacaa
      841 gcagggaaca cccaagtaga agctggctgt cagatcctta tgacggaggc taacaagcag
      901 ggaacaccca agtagaagct ggctgtcaga tctgtcgaat cgaattcctg cagcccggcg
      961 cgcccgggcg atcgcgggta ccgagctcca ggaacatcca aactgagcag ccggggtccc
     1021 ccccaccccc caccccgccc ctcccggcaa ctttgagcct gtgctgggac agagcctcta
     1081 gttcctaaat tagtccatga ggtcagaggc agcactgcca ttgtaacgcg attggagagg
     1141 atcacgtcac cggacacgcc cccaggcatc tccctgggtc tcctaaactt ggcggggaga
     1201 agttttagcc cttaagtttt agcctttaac ccccatattc agaactgtgc gagttggcga
     1261 aaccccacaa atcacaacaa actgtacaca acaccgagct agaggtgatc tttcttgtcc
     1321 attccacaca ggccttagta atgcgtcgcc atagcaacag tgtcactagt agcaccagca
     1381 cttccccaca ccctccccct caggaatccg tactctccag tgaaccccag aaacctctgg
     1441 agagttctgg acaagggcgg aacccacaac tccgattact caagggaggc ggggaagctc
     1501 caccagacgc gaaactgctg gaagattcct ggccccaagg cctcctccgg ctcgctgatt
     1561 ggcccagcgg agagtgggcg gggccggtga agactcctta aaggcgcagg gcggcgagca
     1621 ggtcaccaga cgctgacagc tactcagaac caaatctggt tccatccaga gacaagcgaa
     1681 gacaagagaa gcagagcgag cggcgcgttc ccgatcctcg gccaggacca gccttcccca
     1741 gagcatccct gccgcggagc gcaaccttcc caggagcatc cctgccgcgg agcgcaactt
     1801 tccccggagc atccacgccg cggagcgcag ccttccagaa gcagagcgcg gcgccatggt
     1861 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1921 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1981 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2041 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2101 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2161 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2221 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2281 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2341 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2401 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2461 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2521 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     2581 ccgcgactct agataagtaa tgatcataat cagccatatc acatctgtag aggttttact
     2641 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2701 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2761 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2821 tgtatcttat catgtctgga tccccgggta ccgagctcgc tagagggaca gccccccccc
     2881 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     2941 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     3001 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     3061 gagcctgcag acacctgggg ggatacgggg aaaaagcttt aggctgaaag agagatttag
     3121 aatgacagaa tcatagaacg gcctgggttg caaaggagca cagtgctcat ccagatccaa
     3181 ccccctgcta tgtgcagggt catcaaccag cagcccaggc tgcccagagc cacatccagc
     3241 ctggccttga atgcctgcag ggatggggca tccacagcct ccttgggcaa cctgttcagt
     3301 gcgtcaccac cctctggggg aaaaactgcc tcctcatatc caacccaaac ctcccctgtc
     3361 tcagtgtaaa gccattcccc cttgtcctat caagggggag tttgctgtga cattgttggt
     3421 ctggggtgac acatgtttgc caattcagtg catcacggag aggcagatct tggggataag
     3481 gaagtgcagg acagcatgga cgtgggacat gcaggtgttg agggctctgg gacactctcc
     3541 aagtcacagc gttcagaaca gccttaagga taagaagata ggatagaagg acaaagagca
     3601 agttaaaacc cagcatggag aggagcacaa aaaggccaca gacactgctg gtccctgtgt
     3661 ctgagcctgc atgtttgatg gtgtctggat gcaagcagaa ggggtggaag agcttgcctg
     3721 gagagataca gctgggtcag taggactggg acaggcagct ggagaattgc catgtagatg
     3781 ttcatacaat cgtcaaatca tgaaggctgg aaaagccctc caagatcccc aagaccaacc
     3841 ccaacccacc caccgtgccc actggccatg tccctcagtg ccacatcccc acagttcttc
     3901 atcacctcca gggacggtga cccccccacc tccgtgggca gctgtgccac tgcagcaccg
     3961 ctctttggag aaggtaaatc ttgctaaatc cagcccgacc ctcccctggc acaacgtaag
     4021 gccattatct ctcatccaac tccaggacgg agtcagtgag gatggggctc tagcgagctc
     4081 gaattcattg cagactttca ctggtaccct gtttccttat ctgacctgct ttaactgggt
     4141 aagcttatga aaagtcttgt gtagaagaga aagggataac agcctgtgct aaatgaggaa
     4201 gctgccttgc ctgttcctgc tcagtggggt ttctggctat actacatcaa ctgagtcagt
     4261 gcatcttgac tagttcccac acctttctct gaacagagag agtaaagggc tcaataagaa
     4321 aaatacagtt tatggtctgt acgttgtatt acacgtcatc ccttactttc taaaaggcat
     4381 cttcactgag aaagacatgg atttctaacc ttacaatctc tacaaagttc agaatacata
     4441 ataatcttga attatgatta actgttagtt ttgaccaggt cttctggcag acaggtcaca
     4501 tgtgttagta tcacttattc ctcaagtgtt gatgttagtg tcagcatatt accgatgttc
     4561 cacaaacatt tcctgaatga ctgttaaact tcctacacat taacgagcct ctgcattttt
     4621 ttccagcttc catctatgat ttaagtaaac tctagttttc cacttcttca tattctctct
     4681 ctagatctca attattgcag taccactgtc caagggcaga ggaggttagc tgggcccagg
     4741 cggagtcaat tctctactcc ccaccctgtg ggtgtgttca gccttgtgag ccagcatcag
     4801 gcttgagcac agcagtgctg agtcatgctg agtcatgctg aggcttaggg tgtgtggcca
     4861 gatgttttca gctgtgagtg atcagtgcta tctgggtctc taggaggaag tccacaggga
     4921 aggtgaaaag aaaataagtt tgctccctga agaaaacatt acttacacca gcattacaat
     4981 gaaaagggga ccctgccttg ctgtgtgaca taacctagaa tattttattt tctagttaaa
     5041 aattacttct catgatcttt attagaagtt ataacaaatc attttttaga aatgtaaagt
     5101 ttatttagac tcatacctag aagcaacaga atcacacatt ttagtgtaca cacacacaca
     5161 cacacgcaca cacacacaca tgacttttgt tcataaacta tgcaattgtt aatgaaatgc
     5221 tatttggaat gggcttgaat tcgtattcta tagtgtcacc taaatccaat tcactggccg
     5281 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5341 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5401 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     5461 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     5521 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5581 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5641 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5701 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5761 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5821 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6841 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6901 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6961 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7021 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7081 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7141 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7201 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7261 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7321 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7381 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7441 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7501 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7561 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7621 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7681 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     7741 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     7801 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7861 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctaa tacgactcac
     7921 tatagggaga caagcttgca tgcctgcagg tcgactctag agggacagcc cccccccaaa
     7981 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
     8041 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
     8101 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     8161 cctgcagaca cctgggggga tacggggaaa aagc
//
