LOCUS       sequence-178085-        5934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            GLI1 CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5934)
  AUTHORS   Jackel C, Nogueira MS, Ehni N, Kraus C, Ranke J, Dohmann M, Noessner
            E, Nelson PJ
  TITLE     A vector platform for the rapid and efficient engineering of stable 
            complex transgenes.
  JOURNAL   Sci Rep. 2016 Oct 3;6:34365. doi: 10.1038/srep34365.
  PUBMED    27694838
REFERENCE   2  (bases 1 to 5934)
  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: "Sci Rep."; 
            date: "2016-10-3"; volume: "6:34365. doi"; pages: " 
            10.1038/srep34365"
COMMENT     Sequence Label: pENTR221-GLI1
FEATURES             Location/Qualifiers
     source          1..5934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4046..4145)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4175..4194)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4176..4194)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4199..4215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4199..4215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4317..4336)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4328..5137
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4395..4414)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5284..5872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5773..5792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttgtacct ctgagacgcc atgttcaact cgatgacccc accaccaatc agtagctatg
      721 gcgagccctg ctgtctccgg cccctcccca gtcagggggc ccccagtgtg gggacagaag
      781 gactgtctgg cccgcccttc tgccaccaag ctaacctcat gtccggcccc cacagttatg
      841 ggccagccag agagaccaac agctgcaccg agggcccact cttttcttct ccccggagtg
      901 cagtcaagtt gaccaagaag cgggcactgt ccatctcacc tctgtcggat gccagcctgg
      961 acctgcagac ggttatccgc acctcaccca gctccctcgt agctttcatc aactcgcgat
     1021 gcacatctcc aggaggctcc tacggtcatc tctccattgg caccatgagc ccatctctgg
     1081 gattcccagc ccagatgaat caccaaaaag ggccctcgcc ttcctttggg gtccagcctt
     1141 gtggtcccca tgactctgcc cggggtggga tgatcccaca tcctcagtcc cggggaccct
     1201 tcccaacttg ccagctgaag tctgagctgg acatgctggt tggcaagtgc cgggaggaac
     1261 ccttggaagg tgatatgtcc agccccaact ccacaggcat acaggatccc ctgttgggga
     1321 tgctggatgg gcgggaggac ctcgagagag aggagaagcg tgagcctgaa tctgtgtatg
     1381 aaactgactg ccgttgggat ggctgcagcc aggaatttga ctcccaagag cagctggtgc
     1441 accacatcaa cagcgagcac atccacgggg agcggaagga gttcgtgtgc cactgggggg
     1501 gctgctccag ggagctgagg cccttcaaag cccagtacat gctggtggtt cacatgcgca
     1561 gacacactgg cgagaagcca cacaagtgca cgtttgaagg gtgccggaag tcatactcac
     1621 gcctcgaaaa cctgaagacg cacctgcggt cacacacggg tgagaagcca tacatgtgtg
     1681 agcacgaggg ctgcagtaaa gccttcagca atgccagtga ccgagccaag caccagaatc
     1741 ggacccattc caatgagaag ccgtatgtat gtaagctccc tggctgcacc aaacgctata
     1801 cagatcctag ctcgctgcga aaacatgtca agacagtgca tggtcctgac gcccatgtga
     1861 ccaaacggca ccgtggggat ggccccctgc ctcgggcacc atccatttct acagtggagc
     1921 ccaagaggga gcgggaagga ggtcccatca gggaggaaag cagactgact gtgccagagg
     1981 gtgccatgaa gccacagcca agccctgggg cccagtcatc ctgcagcagt gaccactccc
     2041 cggcagggag tgcagccaat acagacagtg gtgtggaaat gactggcaat gcagggggca
     2101 gcactgaaga cctctccagc ttggacgagg gaccttgcat tgctggcact ggtctgtcca
     2161 ctcttcgccg ccttgagaac ctcaggctgg accagctaca tcaactccgg ccaataggga
     2221 cccggggtct caaactgccc agcttgtccc acaccggtac cactgtgtcc cgccgcgtgg
     2281 gccccccagt ctctcttgaa cgccgcagca gcagctccag cagcatcagc tctgcctata
     2341 ctgtcagccg ccgctcctcc ctggcctctc ctttcccccc tggctcccca ccagagaatg
     2401 gagcatcctc cctgcctggc cttatgcctg cccagcacta cctgcttcgg gcaagatatg
     2461 cttcagccag agggggtggt acttcgccca ctgcagcatc cagcctggat cggataggtg
     2521 gtcttcccat gcctccttgg agaagccgag ccgagtatcc aggatacaac cccaatgcag
     2581 gggtcacccg gagggccagt gacccagccc aggctgctga ccgtcctgct ccagctagag
     2641 tccagaggtt caagagcctg ggctgtgtcc ataccccacc cactgtggca gggggaggac
     2701 agaactttga tccttacctc ccaacctctg tctactcacc acagcccccc agcatcactg
     2761 agaatgctgc catggatgct agagggctac aggaagagcc agaagttggg acctccatgg
     2821 tgggcagtgg tctgaacccc tatatggact tcccacctac tgatactctg ggatatgggg
     2881 gacctgaagg ggcagcagct gagccttatg gagcgagggg tccaggctct ctgcctcttg
     2941 ggcctggtcc acccaccaac tatggcccca acccctgtcc ccagcaggcc tcatatcctg
     3001 accccaccca agaaacatgg ggtgagttcc cttcccactc tgggctgtac ccaggcccca
     3061 aggctctagg tggaacctac agccagtgtc ctcgacttga acattatgga caagtgcaag
     3121 tcaagccaga acaggggtgc ccagtggggt ctgactccac aggactggca ccctgcctca
     3181 atgcccaccc cagtgagggg cccccacatc cacagcctct cttttcccat tacccccagc
     3241 cctctcctcc ccaatatctc cagtcaggcc cctataccca gccaccccct gattatcttc
     3301 cttcagaacc caggccttgc ctggactttg attcccccac ccattccaca gggcagctca
     3361 aggctcagct tgtgtgtaat tatgttcaat ctcaacagga gctactgtgg gagggtgggg
     3421 gcagggaaga tgcccccgcc caggaacctt cctaccagag tcccaagttt ctggggggtt
     3481 cccaggttag cccaagccgt gctaaagctc cagtgaacac atatggacct ggctttggac
     3541 ccaacttgcc caatcacaag tcaggttcct atcccacccc ttcaccatgc catgaaaatt
     3601 ttgtagtggg ggcaaatagg gcttcacata gggcagcagc accacctcga cttctgcccc
     3661 cattgcccac ttgctatggg cctctcaaag tgggaggcac aaaccccagc tgtggtcatc
     3721 ctgaggtggg caggctagga gggggtcctg ccttgtaccc tcctcccgaa ggacaggtat
     3781 gtaaccccct ggactctctt gatcttgaca acactcagct ggactttgtg gctattctgg
     3841 atgagcccca ggggctgagt cctcctcctt cccatgatca gcggggcagc tctggacata
     3901 ccccacctcc ctctgggccc cccaacatgg ctgtgggcaa catgagtgtc ttactgagat
     3961 ccctacctgg ggaaacagaa ttcctcaact ctagtgccta aagagtaggg aatctcatcc
     4021 atcacagatc gcatttccta agggaaccca gctttcttgt acaaagttgg cattataaga
     4081 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     4141 atttgccatc cagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt
     4201 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     4261 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     4321 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     4381 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     4441 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     4501 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     4561 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     4621 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     4681 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     4741 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     4801 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     4861 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     4921 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     4981 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     5041 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     5101 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     5161 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     5221 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5881 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
