LOCUS       sequence-8525-ex        6736 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 6736)
  AUTHORS   Angers S, Thorpe CJ, Biechele TL, Goldenberg SJ, Zheng N, MacCoss 
            MJ, Moon RT
  TITLE     The KLHL12-Cullin-3 ubiquitin ligase negatively regulates the 
            Wnt-beta-catenin pathway by targeting Dishevelled for degradation.
  JOURNAL   Nat Cell Biol. 2006 Apr . 8(4):348-57.
  PUBMED    16547521
REFERENCE   2  (bases 1 to 6736)
  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: "Nat Cell 
            Biol. 2006 Apr . 8(4):348-57."
COMMENT     Sequence Label: XE248 pCDNA3.1+zeo-VSV-KLHL12
FEATURES             Location/Qualifiers
     source          1..6736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             944..976
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     primer_bind     complement(2743..2760)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2749..2973
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3019..3447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3106..3125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3316..3337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3456..3476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3461..3791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3642..3777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3704..3723
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3839..3886
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3905..4279
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4409..4530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4446..4465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4500..4519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4579..4595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4579..4595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4592..4614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4603..4619
                     /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(4627..4633,4634..4651,4652..4657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4672..4693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4810..4827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4981..5569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5061..5080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5740..6531,6532..6600))
                     /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     6363..6382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6601..6705)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgtacaccg acatcgagat
      961 gaaccggctg ggcaagggcg ggggcgcgcc aatgggaggc attatggccc ccaaagacat
     1021 aatgacaaat actcatgcta aatccatcct caattcaatg aactccctca ggaagagcaa
     1081 taccctctgt gatgtgacat tgagagtaga gcagaaagac ttccctgccc atcggattgt
     1141 gctggctgcc tgtagtgatt acttctgtgc catgttcact agtgagctct cagagaaggg
     1201 gaaaccttat gttgacatcc aaggtttgac tgcctctacc atggaaattt tattggactt
     1261 tgtgtacaca gaaacggtac atgtgacagt ggagaatgta caagaactgc ttcctgcagc
     1321 ctgtctgctt cagttgaaag gtgtgaaaca agcctgctgt gagttcttag aaagtcagtt
     1381 ggacccttct aattgcctgg gtattaggga ttttgctgaa acccacaatt gtgttgacct
     1441 gatgcaagca gctgaggttt ttagccagaa gcattttcct gaagtggtac agcatgaaga
     1501 gttcattctt ctgagtcaag gagaggtgga aaagctaatc aagtgcgacg aaattcaggt
     1561 ggattctgaa gagccagtct ttgaggctgt catcaactgg gtgaagcatg ccaagaaaga
     1621 gcgggaagaa tccttgccta acctgctaca gtatgtgcgg atgcccctac taacccccag
     1681 gtatatcaca gatgtaatag atgctgagcc tttcatccgc tgtagtttac aatgcaggga
     1741 tctggttgat gaagcaaaga agtttcatct gaggcctgaa cttcggagtc agatgcaggg
     1801 acccaggaca agggctcgcc taggagccaa tgaagtgctt ttggtggttg ggggctttgg
     1861 aagccagcag tctcccattg atgtggtaga gaaatatgac cccaagactc aggagtggag
     1921 ctttttgcca agcatcactc gtaagagacg ttatgtggcc tcagtgtccc ttcatgaccg
     1981 gatctacgtc attggtggct atgatggccg ttcccgcctt agttcagtgg aatgtctaga
     2041 ctacacagca gatgaggatg gggtctggta ttctgtggcc cctatgaatg tccgacgagg
     2101 tcttgctgga gccaccaccc tgggagatat gatctatgtc tctggaggct ttgatggaag
     2161 caggcgtcac accagtatgg agcgctatga tccaaacatt gaccagtgga gcatgctggg
     2221 ggatatgcag acagcccggg aaggtgccgg actcgtagtg gccagtggag tgatctactg
     2281 tctaggagga tatgacggct tgaatatctt aaattcagtt gagaaatacg accctcatac
     2341 aggacattgg actaatgtta caccaatggc caccaagcgt tctggtgcag gagtagccct
     2401 gctgaatgac catatttatg tggtgggggg atttgatggt acagcccacc tttcttccgt
     2461 tgaagcatac aacattcgca ctgattcctg gacaactgtc accagtatga ccactccacg
     2521 atgctatgta ggggccacag tgcttcgggg gagactctat gcaattgcag gatatgatgg
     2581 taattccctg ctaagtagca ttgaatgtta tgaccctatc atcgacagct gggaagtcgt
     2641 gacatccatg ggaacccagc gctgtgatgc tggtgtttgt gttctccgcg agaagtgagc
     2701 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     2761 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2821 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2881 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2941 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3001 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     3181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3361 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     3421 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3481 aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3661 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3721 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3781 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     3841 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     3901 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     3961 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     4021 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     4081 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     4141 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     4201 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     4261 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     4321 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4381 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4441 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4501 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4561 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4621 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4681 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4741 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5161 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5401 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5821 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5881 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5941 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6001 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6061 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6121 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6181 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6241 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6301 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6361 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6421 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6481 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6541 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6721 agtgccacct gacgtc
//
