LOCUS       sequence-193940-        6554 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 6554)
  AUTHORS   Kinzler KW, Ruppert JM, Bigner SH, Vogelstein B
  TITLE     The GLI gene is a member of the Kruppel family of zinc finger 
            proteins.
  JOURNAL   Nature. 1988 Mar 24. 332(6162):371-4.
  PUBMED    2832761
REFERENCE   2  (bases 1 to 6554)
  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: "Nature. 
            1988 Mar 24. 332(6162):371-4."
COMMENT     Sequence Label: GLI K12
FEATURES             Location/Qualifiers
     source          1..6554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(128..716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(208..227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(887..1678,1679..1747))
                     /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     1510..1529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1748..1852)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1878..2333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2015..2036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2227..2246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2460..2482
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2474..2491
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2475..2491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2498..2517
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2498..2516
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2549..2565
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2549..2565
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6196..6212)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6197..6213)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6241..6259)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6280..6296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6280..6296)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6293..6315)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6304..6320
                     /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(6328..6334,6335..6352,6353..6358))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6373..6394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6511..6528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
       61 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1861 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1921 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1981 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2041 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2101 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2161 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2221 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2281 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     2341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2461 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
     2521 attggagctc caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga
     2581 attccccttt attatcagga aacagtgtca ttatcaaatt tctcctcaaa acgtatacag
     2641 actatacata gcccccagcc catacctccc atccctgggg catcttgtgc aggggactgc
     2701 agctccccca atttttctgg aaggatagaa accccttagg aaatgcgatc tgtgatggat
     2761 gagattccct actctttagg cactagagtt gaggaattct gtttccccag gtagggatct
     2821 cagtaagaca ctcatgttgc ccacagccat gttggggggc ccagagggag gtggggtatg
     2881 tccagagctg ccccgctgat catgggaagg aggaggactc agcccctggg gctcatccag
     2941 aatagccaca aagtccagct gagtgttgtc aagatcaaga gagtccaggg ggttacatac
     3001 ctgtccttcg ggaggagggt acaaggcagg accccctcct agcctgccca cctcaggatg
     3061 accacagctg gggtttgtgc ctcccacttt gagaggccca tagcaagtgg gcaatggggg
     3121 cagaagtcga ggtggtgctg ctgccctatg tgaagcccta tttgccccca ctacaaaatt
     3181 ttcatggcat ggtgaagggg tgggatagga acctgacttg tgattgggca agttgggtcc
     3241 aaagccaggt ccatatgtgt tcactggagc tttagcacgg cttgggctaa cctgggaacc
     3301 ccccagaaac ttgggactct ggtaggaagg ttcctgggcg ggggcatctt ccctgccccc
     3361 accctcccac agtagctcct gttgagattg aacataatta cacacaagct gagccttgag
     3421 ctgccctgtg gaatgggtgg gggaatcaaa gtccaggcaa ggcctgggtt ctgaaggaag
     3481 ataatcaggg ggtggctggg tataggggcc tgactggaga tattggggag gagagggctg
     3541 ggggtaatgg gaaaagagag gctgtggatg tgggggcccc tcactggggt gggcattgag
     3601 gcagggtgcc agtcctgtgg agtcagaccc cactgggcac ccctgttctg gcttgacttg
     3661 cacttgtcca taatgttcaa gtcgaggaca ctggctgtag gttccaccta gagccttggg
     3721 gcctgggtac agcccagagt gggaagggaa ctcaccccat gtttcttggg tggggtcagg
     3781 atatgaggcc tgctggggac aggggttggg gccatagttg gtgggtggac caggcccaag
     3841 aggcagagag cctggacccc tcgctccata aggctcagct gctgcccctt caggtccccc
     3901 atatcccaga gtatcagtag gtgggaagtc catatagggg ttcagaccac tgcccaccat
     3961 ggaggtccca acttctggct cttcctgtag ccctctagca tccatggcag cattctcagt
     4021 gatgctgggg ggctgtggtg agtagacaga ggttgggagg taaggatcaa agttctgtcc
     4081 tccccctgcc acagtgggtg gggtatggac acagcccagg ctcttgaacc tctggactct
     4141 agctggagca ggacggtcag cagcctgggc tgggtcactg gccctccggg tgacccctgc
     4201 attggggttg tatcctggat actcggctcg gcttctccaa ggaggcatgg gaagaccacc
     4261 tatccgatcc aggctggatg ctgcagtggg cgaagtacca ccccctctgg ctgaagcata
     4321 tcttgcccga agcaggtagt gctgggcagg cataaggcca ggcagggagg atgctccatt
     4381 ctctggtggg gagccagggg ggaaaggaga ggccagggag gagcggcggc tgacagtata
     4441 ggcagagctg atgctgctgg agctgctgct gcggcgttca agagagactg gggggcccac
     4501 gcggcgggac acagtggtac cggtgtggga caagctgggc agtttgagac cccgggtccc
     4561 tattggccgg agttgatgta gctggtccag cctgaggttc tcaaggcggc gaagagtgga
     4621 cagaccagtg ccagcaatgc aaggtccctc gtccaagctg gagaggtctt cagtgctgcc
     4681 ccctgcattg ccagtcattt ccacaccact gtctgtattg gctgcactcc ctgccgggga
     4741 gtggtcactg ctgcaggatg actgggcccc agggcttggc tgtggcttca tggcaccctc
     4801 tggcacagtc agtctgcttt cctccctgat gggacctcct tcccgctccc tcttgggctc
     4861 cactgtagaa atggatggtg cccgaggcag ggggccatcc ccacggtgcc gtttggtcac
     4921 atgggcgtca ggaccatgca ctgtcttgac atgttttcgc agcgagctag gatctgtata
     4981 gcgtttggtg cagccaggga gcttacatac atacggcttc tcattggaat gggtccgatt
     5041 ctggtgcttg gctcggtcac tggcattgct gaaggcttta ctgcagccct cgtgctcaca
     5101 catgtatggc ttctcacccg tgtgtgaccg caggtgcgtc ttcaggtttt cgaggcgtga
     5161 gtatgacttc cggcaccctt caaacgtgca cttgtgtggc ttctcgccag tgtgtctgcg
     5221 catgtgaacc accagcatgt actgggcttt gaagggcctc agctccctgg agcagccccc
     5281 ccagtggcac acgaactcct tccgctcccc gtggatgtgc tcgctgttga tgtggtgcac
     5341 cagctgctct tgggagtcaa attcctggct gcagccatcc caacggcagt cagtttcata
     5401 cacagattca ggctcacgct tctcctctct ctcgaggtcc tcccgcccat ccagcatccc
     5461 caacagggga tcctgtatgc ctgtggagtt ggggctggac atatcacctt ccaagggttc
     5521 ctcccggcac ttgccaacca gcatgtccag ctcagacttc agctggcaag ttgggaaggg
     5581 tccccgggac tgaggatgtg ggatcatccc accccgggca gagtcatggg gaccacaagg
     5641 ctggacccca aaggaaggcg agggcccttt ttggtgattc atctgggctg ggaatcccag
     5701 agatgggctc atggtgccaa tggagagatg accgtaggag cctcctggag atgtgcatcg
     5761 cgagttgatg aaagctacga gggagctggg tgaggtgcgg ataaccgtct gcaggtccag
     5821 gctggcatcc gacagaggtg agatggacag tgcccgcttc ttggtcaact tgactgcact
     5881 ccggggagaa gaaaagagtg ggccctcggt gcagctgttg gtctctctgg ctggcccata
     5941 actgtggggg ccggacatga ggttagcttg gtggcagaag ggcgggccag acagtccttc
     6001 tgtccccaca ctgggggccc cctgactggg gaggggccgg agacagcagg gctcgccata
     6061 gctactgatt ggtggtgggg tcatcgagtt gaacatggcg tctcagagga gggtgtgggg
     6121 acactctgtc tgggcgctgg gctcgggatg cgcggtccag ggctggagtc tgggggaatt
     6181 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtacca gcttttgttc
     6241 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6541 tcggctgcgg cgag
//
