LOCUS       sequence-241756-        5315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gli activity reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5315)
  AUTHORS   Sasaki H, Hui C, Nakafuku M, Kondoh H
  TITLE     A binding site for Gli proteins is essential for HNF-3beta floor 
            plate enhancer activity in transgenics and can respond to Shh in 
            vitro.
  JOURNAL   Development. 1997 Apr . 124(7):1313-22.
  PUBMED    9118802
REFERENCE   2  (bases 1 to 5315)
  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: 
            "Development. 1997 Apr . 124(7):1313-22."
COMMENT     Sequence Label: GBSGFP in pBS-KS
FEATURES             Location/Qualifiers
     source          1..5315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     61..77
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     62..78
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1470..1472,1473..1475,1476..2189)
                     /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(1470..1472,1473..1475,1476..2186)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1515..1536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1776..1795)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2123..2144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2322..2456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2359..2378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2413..2432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2453..2469)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2453..2469)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2504..2523)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2505..2523)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2530..2547)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2530..2546)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2539..2561)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2687..3142)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2824..2845)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3036..3055
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3169..3273
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3274..3342,3343..4134)
                     /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(3492..3511)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4305..4893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4794..4813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5047..5064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5181..5202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5217..5222,5223..5240,5241..5247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5255..5271
                     /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     5260..5282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5279..5295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5279..5295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga agcggccgct
       61 tcgaggtcga cggtatcgat aagcttgatt cgaggatcct tatgacggag gctaacaagc
      121 agggaacacc caagtagaag ctggctgtca gatccttatg acggaggcta acaagcaggg
      181 aacacccaag tagaagctgg ctgtcagatc cttatgacgg aggctaacaa gcagggaaca
      241 cccaagtaga agctggctgt cagatcctta tgacggaggc taacaagcag ggaacaccca
      301 agtagaagct ggctgtcaga tccttatgac ggaggctaac aagcagggaa cacccaagta
      361 gaagctggct gtcagatcct tatgacggag gctaacaagc agggaacacc caagtagaag
      421 ctggctgtca gatccttatg acggaggcta acaagcaggg aacacccaag tagaagctgg
      481 ctgtcagatc cttatgacgg aggctaacaa gcagggaaca cccaagtaga agctggctgt
      541 cagatctgtc gaatcgaatt cctgcagccc ggcgcgcccg ggcgatcgcg ggtaccgagc
      601 tccaggaaca tccaaactga gcagccgggg tcccccccac cccccacccc gcccctcccg
      661 gcaactttga gcctgtgctg ggacagagcc tctagttcct aaattagtcc atgaggtcag
      721 aggcagcact gccattgtaa cgcgattgga gaggatcacg tcaccggaca cgcccccagg
      781 catctccctg ggtctcctaa acttggcggg gagaagtttt agcccttaag ttttagcctt
      841 taacccccat attcagaact gtgcgagttg gcgaaacccc acaaatcaca acaaactgta
      901 cacaacaccg agctagaggt gatctttctt gtccattcca cacaggcctt agtaatgcgt
      961 cgccatagca acagtgtcac tagtagcacc agcacttccc cacaccctcc ccctcaggaa
     1021 tccgtactct ccagtgaacc ccagaaacct ctggagagtt ctggacaagg gcggaaccca
     1081 caactccgat tactcaaggg aggcggggaa gctccaccag acgcgaaact gctggaagat
     1141 tcctggcccc aaggcctcct ccggctcgct gattggccca gcggagagtg ggcggggccg
     1201 gtgaagactc cttaaaggcg cagggcggcg agcaggtcac cagacgctga cagctactca
     1261 gaaccaaatc tggttccatc cagagacaag cgaagacaag agaagcagag cgagcggcgc
     1321 gttcccgatc ctcggccagg accagccttc cccagagcat ccctgccgcg gagcgcaacc
     1381 ttcccaggag catccctgcc gcggagcgca actttccccg gagcatccac gccgcggagc
     1441 gcagccttcc agaagcagag cgcggcgcca tggtgagcaa gggcgaggag ctgttcaccg
     1501 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1561 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1621 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1681 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1741 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1801 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1861 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1921 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1981 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2041 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     2101 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2161 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagataa gtaatgatca
     2221 taatcagcca tatcacatct gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2281 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2341 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2401 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccact
     2461 agttctagag cggccgccac cgcggtgcgt cgacgccaat tcgccctata gtgagtcgta
     2521 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2581 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2641 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt
     2701 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2761 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2821 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2881 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2941 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3001 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3061 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3121 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3181 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3241 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3301 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3361 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3421 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3481 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3541 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3601 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3661 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3721 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3781 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3841 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3901 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3961 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4021 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4081 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4141 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4441 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4981 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5041 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     5101 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     5161 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     5221 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     5281 ggaaacagct atgaccatga ttacgccaag ctcga
//
