LOCUS       sequence-194136-        9096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9096)
  AUTHORS   Roessler E, Ermilov AN, Grange DK, Wang A, Grachtchouk M, Dlugosz 
            AA, Muenke M
  TITLE     A previously unidentified amino-terminal domain regulates 
            transcriptional activity of wild-type and disease-associated human 
            GLI2.
  JOURNAL   Hum Mol Genet. 2005 Aug 1. 14(15):2181-8.
  PUBMED    15994174
REFERENCE   2  (bases 1 to 9096)
  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: "Hum Mol 
            Genet. 2005 Aug 1. 14(15):2181-8."
COMMENT     Sequence Label: pCS2-MT GLI2 FL
FEATURES             Location/Qualifiers
     source          1..9096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1071
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1106..1124
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1106..1123
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1174..1203
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1213..1242
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1252..1281
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1291..1320
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1330..1359
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1393..1422
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    6219..6353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6243..6262)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6297..6316
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6466..6486)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6466..6484)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6505..6521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6505..6521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6518..6540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6529..6545
                     /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)."
     promoter        complement(join(6553..6559,6560..6577,6578..6583))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6598..6619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6736..6753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6907..7495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6987..7006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7666..8457,8458..8526))
                     /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     8289..8308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8527..8631)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(8657..9096,1..16))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8794..8815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9006..9025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
       61 gtgcgggcct cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact
      121 catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact
      181 cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac
      241 catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca
      301 cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca
      361 tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc
      421 aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg
      481 ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg
      541 gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt
      601 caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat
      661 atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt
      721 gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc
      781 cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa
      841 tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg
      901 cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc
      961 cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc
     1021 ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa ccgccattct
     1081 gcctggggac gtcggagcaa gcttgattta ggtgacacta tagaatacaa gctacttgtt
     1141 ctttttgcag gatcccatcg atttaaagct atggagcaaa agctcatttc tgaagaggac
     1201 ttgaatgaaa tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag
     1261 ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga agaggacttg
     1321 aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga gagcttgggc
     1381 gacctcacca tggagcaaaa gctcatttct gaagaggact tgaattcaat ggagacgtct
     1441 gcctcagcca ctgcctccga gaagcaagaa gccaaaagtg ggatcctgga ggccgctggc
     1501 ttccccgacc cgggtaaaaa ggcctctcct ttggtggtgg ctgcagcggc agcagcagcg
     1561 gtagctgccc aaggagtgcc gcagcatctc ttgccaccat tccatgcgcc cctaccgatt
     1621 gacatgcgac accaggaagg aaggtaccat tacgagcctc attctgtcca cggtgtgcac
     1681 gggccccctg ccctcagcgg cagccctgtc atctctgaca tctccttgat ccggctttcc
     1741 ccgcacccgg ctggccctgg ggagtccccc ttcaacgccc cccacccgta cgtgaacccc
     1801 cacatggagc actacctccg ttctgtgcac agcagcccca cgctctccat gatctctgca
     1861 gccaggggcc tcagccccgc tgatgtggcc caggagcacc ttaaggagag gggactgttt
     1921 ggccttcctg ctccaggcac caccccctca gactattacc accagatgac cctcgtggca
     1981 ggccaccccg cgccctacgg ggacctgctg atgcagagcg ggggcgctgc cagcgcaccc
     2041 catctccacg actacctcaa ccccgtggac gtgtcccgtt tctccagccc gcgggtgacg
     2101 ccccgcctga gccgcaagcg ggcgctgtcc atctccccac tctcagacgc cagcctggac
     2161 ctgcagcgga tgatctgcac ctcacccaac tcgctagtgg cctacatcaa caactcccga
     2221 agcagctcag cggccagcgg ttcctacggg catctgtcag cgggtgccct cagcccagcc
     2281 ttcaccttcc cccaccccat caaccccgtg gcctaccagc agattctgag ccagcagagg
     2341 ggtctggggt cagcctttgg acacacacca cccctgatcc agccctcacc caccttcctg
     2401 gcccagcagc ccatggccct cacctccatc aatgccacgc ccacccagct cagcagcagc
     2461 agcaactgtc tgagtgacac caaccagaac aagcagagca gtgagtcggc cgtcagcagc
     2521 accgtcaacc ctgtcgccat tcacaagcgc agcaaggtca agaccgagcc tgagggcctg
     2581 cggccggcct cccctctggc gctgacgcag ggccaggtgt ctggacacgg ctcatgtggg
     2641 tgtgcccttc ccctctccca ggagcagctg gctgacctca aggaagatct ggacagggat
     2701 gactgtaagc aggaggctga ggtggtcatc tatgagacca actgccactg ggaagactgc
     2761 accaaggagt acgacaccca ggagcagctg gtgcatcaca tcaacaacga gcacatccac
     2821 ggggagaaga aggagtttgt gtgccgctgg caggcctgca cgcgggagca gaagcccttc
     2881 aaggcgcagt acatgctggt ggtgcacatg cggcgacaca cgggcgagaa gccccacaag
     2941 tgcacgttcg agggctgctc gaaggcctac tcccgcctgg agaacctgaa gacacacctg
     3001 cggtcccaca ccggggagaa gccatatgtg tgtgagcacg agggctgcaa caaagccttc
     3061 tccaacgcct cggaccgcgc caagcaccag aatcgcaccc actccaacga gaaaccctac
     3121 atctgcaaga tcccaggctg caccaagaga tacacagacc ccagctctct ccggaagcat
     3181 gtgaaaacgg tccacggccc agatgcccac gtcaccaaga agcagcgcaa tgacgtgcac
     3241 ctccgcacac cgctgctcaa agagaatggg gacagtgagg ccggcacgga gcctggcggc
     3301 ccagagagca ccgaggccag cagcaccagc caggccgtgg aggactgcct gcacgtcaga
     3361 gccatcaaga ccgagagctc cgggctgtgt cagtccagcc ccggggccca gtcgtcctgc
     3421 agcagcgagc cctctcctct gggcagtgcc cccaacaatg acagtggcgt ggagatgccg
     3481 gggacggggc ccgggagcct gggagacctg acggcactgg atgacacacc cccaggggcc
     3541 gacacctcag ccctggctgc cccctccgct ggtggcctcc agctgcgcaa acacatgacc
     3601 accatgcacc ggttcgagca gctcaagaag gagaagctca agtcactcaa ggattcctgc
     3661 tcatgggccg ggccgactcc acacacgcgg aacaccaagc tgcctcccct cccgggaagt
     3721 ggctccatcc tggaaaactt cagtggcagt gggggcggcg ggcccgcggg gctgctgccg
     3781 aacccgcggc tgtcggagct gtccgcgagc gaggtgacca tgctgagcca gctgcaggag
     3841 cgccgcgaca gctccaccag cacggtcagc tcggcctaca ccgtgagccg ccgctcctcc
     3901 ggcatctccc cctacttctc cagccgccgc tccagcgagg cctcgcccct gggcgccggc
     3961 cgcccgcaca acgcgagctc cgctgactcc tacgacccca tctccacgga cgcgtcgcgg
     4021 cgctcgagcg aggccagcca gtgcagcggc ggctccgggc tgctcaacct cacgccggcg
     4081 cagcagtaca gcctgcgggc caagtacgcg gcagccactg gcggcccccc gcccactccg
     4141 ctgccgggcc tggagcgcat gagcctgcgg accaggctgg cgctgctgga cgcgcccgag
     4201 cgcacgctgc ccgccggctg cccacgccca ctggggccgc ggcgtggcag cgacgggccg
     4261 acctatggcc acggccacgc gggggctgcg cccgccttcc cccacgaggc tccaggcggc
     4321 ggagccaggc gggccagcga ccctgtgcgg cggcccgatg ccctgtccct gccgcgggtg
     4381 cagcgcttcc acagcaccca caacgtgaac cccggcccgc tgccgccctg tgccgacagg
     4441 cgaggcctcc gcctgcagag ccacccgagc accgacggcg gcctggcccg cggcgcctac
     4501 tcgccccggc cgcctagcat cagcgagaac gtggcgatgg aggccgtggc ggcaggagtg
     4561 gacggcgcgg ggcccgaggc cgacctgggg ctgccggagg acgacctggt gcttccagac
     4621 gacgtggtgc agtacatcaa ggcgcacgcc agtggcgctc tggacgaggg caccgggcag
     4681 gtgtatccca cggaaagcac tggcttctct gacaacccca gactacccag cccggggctg
     4741 cacggccagc gcaggatggt ggctgcggac tccaacgtgg gcccctccgc ccctatgctg
     4801 ggaggatgcc agttaggctt tggggcgccc tccagcctga acaaaaataa catgcctgtg
     4861 cagtggaatg aggtgagctc cggcaccgta gacgccctgg ccagccaggt gaagcctcca
     4921 ccctttcctc agggcaacct ggcggtggtg cagcagaagc ctgcctttgg ccagtacccg
     4981 ggctacagtc cgcaaggcct acaggctagc cctgggggcc tggacagcac gcagccacac
     5041 ctgcagcccc gcagcggagc cccctcccag ggcatcccca gggtaaacta catgcagcag
     5101 ctgcgacagc cagtggcagg cagccagtgt cctggcatga ctaccactat gagcccccat
     5161 gcctgctatg gccaagtcca cccccagctg agccccagca ccatcagtgg ggccctcaac
     5221 cagttccccc aatcctgcag caacatgcca gccaagccag ggcatctggg gcaccctcag
     5281 cagacagaag tggcacctga ccccaccacg atgggcaatc gccacaggga acttggggtc
     5341 cccgattcag ccctggctgg agtgccacca cctcacccag tccagagcta cccacagcag
     5401 agccatcacc tggcagcctc catgagccag gagggctacc accaggtccc cagccttctg
     5461 cctgcccgcc agcctggctt catggagccc caaacaggcc cgatgggggt ggctacagca
     5521 ggctttggcc tagtgcagcc ccggcctccc ctcgagccca gccccactgg ccgccaccgt
     5581 ggggtacgtg ctgtgcagca gcagctggcc tacgccaggg ccacaggcca tgccatggct
     5641 gccatgccgt ccagtcagga aacagcagag gctgtgccca agggagcgat gggcaacatg
     5701 gggtcggtgc ctccccagcc gcctccgcag gacgcaggtg gggccccgga ccacagcatg
     5761 ctctactact acggccagat ccacatgtac gaacaggatg gaggcctgga gaacctcggg
     5821 agctgccagg tcatgcggtc ccagccacca cagccacagg cctgtcagga cagcatccag
     5881 ccccagccct tgccctcacc aggggtcaac caggtgtcca gcactgtgga ctcccagctc
     5941 ctggaggccc cccagattga cttcgatgcc atcatggatg atggcgatca ctcgagtttg
     6001 ttctcgggtg ctctgagccc cagcctcctc cacagcctct cccagaactc ctcccgcctc
     6061 accacccccc gaaactcctt gaccctgccc tccatccccg caggcatcag caacatggct
     6121 gtcggggaca tgagctccat gctcaccagc ctcgccgagg agagcaagtt cctgaacatg
     6181 atgacctagt ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     6241 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     6301 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     6361 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     6421 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     6481 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6541 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     6601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6661 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6721 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7321 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8521 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8581 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8641 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     8701 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     8761 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     8821 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     8881 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     8941 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     9001 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     9061 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcg
//
