LOCUS       sequence-43806-e        6922 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 6922)
  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 6922)
  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: XE253 pCDNA3.1(zeo)-hDsh3-82-716
FEATURES             Location/Qualifiers
     source          1..6922
                     /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"
     regulatory      935..944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             944..967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1310..1321
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     primer_bind     complement(2929..2946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2935..3159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3205..3633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3292..3311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3502..3523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3642..3662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3647..3977
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3828..3963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3890..3909
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4025..4072
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4091..4465
                     /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    4595..4716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4632..4651)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4686..4705
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4765..4781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4765..4781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4778..4800)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4789..4805
                     /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(4813..4819,4820..4837,4838..4843))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4858..4879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4996..5013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5167..5755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5247..5266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5926..6717,6718..6786))
                     /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     6549..6568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6787..6891)
                     /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 atggactaca aggacgacga
      961 cgacaagggc ggcgcgccag gctcacaccc agacccagcc cccttctgtg ctgataaccc
     1021 atcggagctg ccaccaccta tggagcgcac gggaggcatc ggggactccc gacccccatc
     1081 cttccaccct catgctggtg ggggcagcca ggagaacctg gacaatgaca cagagacgga
     1141 ctctttggtg tctgcccagc gagagcggcc acgccggagg gatggcccag agcatgcaac
     1201 ccggctaaat ggaactgcga agggggaacg gcggcgagaa ccagggggtt atgatagctc
     1261 atccaccctt atgagcagtg agctggagac caccagcttc tttgactcag atgaggatga
     1321 ctccaccagc aggttcagca gctccacaga acagagcagt gcctcacgcc tgatgagaag
     1381 acacaagcgg cggcggcgga agcagaaggt ttctcggatt gagcggtcct cgtccttcag
     1441 cagcatcacg gactccacca tgtcactcaa catcatcacg gtcactctca acatggaaaa
     1501 atataacttc ttgggcatct ccattgtggg ccaaagcaac gagcgtggtg acggcggcat
     1561 ctacattggc tctatcatga agggtggggc cgtggctgct gatggacgca tcgagccagg
     1621 agatatgttg ttacaggtaa acgagatcaa ctttgagaac atgagtaatg acgatgcagt
     1681 ccgggtactg cgggagattg tgcacaaacc ggggcccatc accctgactg tagccaagtg
     1741 ctgggaccca agtccacgtg gttgcttcac attgcccagg agcgagccca tccggcccat
     1801 tgaccctgcg gcctgggtct cccacactgc agccatgacc ggcaccttcc ctgcatacgg
     1861 catgagcccc tccctgagca ccatcacctc caccagctcc tccatcacca gttccatccc
     1921 tgacacagag cgcctagacg acttccactt gtccatccac agtgacatgg ctgccatcgt
     1981 aaaagccatg gcctcccctg aatcagggtt ggaggtccgt gaccgcatgt ggctcaagat
     2041 taccatccct aatgctttca tcggctcaga tgtggtggac tggctgtacc acaatgtgga
     2101 aggcttcacg gaccggaggg aggcccgcaa gtatgccagc aacctgctga aagctggctt
     2161 catccgccat accgtcaaca agatcacctt ctccgagcag tgctactaca tcttcggtga
     2221 cctctgcggc aacatggcca acctgtctct ccacgatcac gatggctcca gtggcgcctc
     2281 tgaccaggac acactggccc ctttgccgca cccgggggcc gccccttggc ccatggcttt
     2341 cccgtaccag tacccgccac ccccgcaccc atacaacccg cacccgggct tcccggagct
     2401 gggctacagc tacggcgggg gcagcgccag cagtcagcac agcgaaggca gtcggagcag
     2461 tggctccaac cgtagcggca gcgatcggag gaaggagaag gacccgaagg ccggggactc
     2521 caagtccggg ggcagcggca gcgaatcgga ccacaccaca cgcagcagcc tgcgggggcc
     2581 gcgggagcgg gcgcccagcg agcgctcagg tccggcggcc agcgagcaca gccaccgcag
     2641 ccaccattcc ctggccagca gccttcgcag ccaccacaca cacccgagct acggtcctcc
     2701 cggagtgccc cctctctacg gcccccccat gctgatgatg cccccgccgc ccgcggccat
     2761 ggggccccca ggagcccctc cgggccgcga cctggcctca gtgcccccgg aactgaccgc
     2821 cagcagacag tccttccgca tggccatggg aaaccccagt gagttctttg tggatgtgat
     2881 gtgagcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     2941 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3001 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3061 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3121 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3181 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3361 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3481 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3541 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     3601 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3661 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3901 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3961 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4021 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4081 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     4141 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     4201 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     4261 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     4321 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     4381 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     4441 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     4501 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4561 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4621 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4681 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4741 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4921 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5341 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6241 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6541 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6901 ccgaaaagtg ccacctgacg tc
//
