LOCUS       sequence-251111-        7298 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 7298)
  AUTHORS   Choi JH, Zhong X, McAlpine W, Liao TC, Zhang D, Fang B, Russell J, 
            Ludwig S, Nair-Gill E, Zhang Z, Wang KW, Misawa T, Zhan X, Choi M, 
            Wang T, Li X, Tang M, Sun Q, Yu L, Murray AR, Moresco EMY, Beutler B
  TITLE     LMBR1L regulates lymphopoiesis through Wnt/beta-catenin signaling.
  JOURNAL   Science. 2019 May 10;364(6440). pii: 364/6440/eaau0812. doi: 
            10.1126/science.aau0812.
  PUBMED    31073040
REFERENCE   2  (bases 1 to 7298)
  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: "Science."; 
            date: "2019-05-10"; volume: "364(6440). pii: 364/6440/eaau0812. 
            doi"; pages: " 10.1126/science.aau0812"
COMMENT     Sequence Label: pcDNA6-N-3XFLAG-Dvl2
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             696..719
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1380..1391
                     /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(3020..3037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3026..3250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3296..3724
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3383..3402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3593..3614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3733..3753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3738..4067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3918..4053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3980..3999
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4115..4162
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4181..4579
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4737..4858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4774..4793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4828..4847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4907..4923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4920..4942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4931..4947
                     /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(4955..4961,4962..4979,4980..4985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5000..5021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5138..5155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5309..5897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5389..5408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6068..6859,6860..6928))
                     /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     6691..6710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6929..7033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7108..7127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaga gctccttaag gttaacgcca ccatggacta caaagacgat gacgacaagt
      721 ctagagaatt cggatccagc gagaccatgg cgggcagcag cgcggggggc ggtggtgtag
      781 gcgagacgaa ggtgatttac catctggatg aagaagagac tccttacctg gtgaagatcc
      841 ctgtcccggc ggagcgcatc acgctcggcg atttcaagag cgttttgcag cggcccgcgg
      901 gcgccaagta ctttttcaag tccatggatc aggattttgg ggtggtgaag gaagagatct
      961 ccgatgacaa tgcccgccta ccttgcttca atggaagggt tgtctcctgg cttgtgtcgt
     1021 cagatacccc acagcctgag gtggctcccc cagcccatga gtctcggaca gaactggttc
     1081 ctccgcctcc acccttaccc cctttgccac cagaaaggac cagtggaatt ggggactcaa
     1141 ggcctccatc cttccaccct aatgtatcca gcagccatga aaatctagag cctgagacag
     1201 agaccgaatc tgtcgtatca ctgaggcgag accgacctag gaggagagac agcagtgaac
     1261 atggcgctgg tggccacagg cctggtggcc cctcgaggct ggagcgccac ctggctggct
     1321 acgagagctc ttccaccctc atgaccagtg agctggagag taccagccta ggagactcgg
     1381 atgaggatga caccatgagt aggttcagca gctccactga gcagagcagt gcctcccgcc
     1441 tcctcaagcg ccaccgaagg cgaaggaagc agcggccgcc acgcatggag aggacctcat
     1501 ccttcagcag tgtcaccgat tccacaatgt ctctcaacat catcacggtc acgctcaaca
     1561 tggagaagta caacttcctg ggcatctcca ttgtgggcca aagtaacgag cgtggtgatg
     1621 ggggcatcta catcggctcc atcatgaaag ggggcgcggt ggctgcggac gggcggatcg
     1681 agcctggaga catgcttttg caggtaaatg atatgaactt tgagaacatg agcaacgacg
     1741 atgctgtacg agtgctgaga gacatcgtgc acaagccagg ccccatcgtg ctgaccgtgg
     1801 ccaagtgttg ggacccgtct ccccaggcct acttcaccct ccctcgaaat gagcccatcc
     1861 agcccatcga cccagccgcc tgggtgtcgc actcggctgc gctgactggc gccttccctg
     1921 cttaccctgg ctcctcatct atgagcacta tcacatctgg ctcctctctg cctgatggct
     1981 gcgaaggccg gggtctctct gtccacatgg acatggcctc tgtcaccaag gccatggcag
     2041 ccccagagtc tgggctcgaa gtccgggacc gcatgtggct caagatcacc atcccaaacg
     2101 cctttctagg ctccgatgtg gtggactggc tgtaccatca cgtggaaggt tttcctgagc
     2161 gccgggaggc ccgcaagtat gccagtgggc tgttgaaggc gggactcatc cggcacaccg
     2221 tcaacaagat tactttctct gagcagtgct attatgtctt cggggacctc agtggtggct
     2281 gtgagagtta cctagttaac ctctctctga atgacaatga cggttccagt ggggcttcag
     2341 accaggatac cctggcacct ctgcctggag ctactccctg gcccctgctg cccaccttct
     2401 cctaccagta tccagcgcca cacccctaca gcccccagcc tccaccctac cacgagcttt
     2461 cttcgtacac ctatggtgga ggcagtgcca gcagccagca cagtgagggg agccggagca
     2521 gcgggtcgac aagaagcgat gggggggctg ggcgcacagg caggcctgag gaacgggccc
     2581 ctgagtccaa gtccggcagt ggcagtgagt cagaactctc cagccgggga ggcagccttc
     2641 ggcggggtgg ggagcctggt gggactggtg atggcggccc tcctccatcc aggggctcga
     2701 caggcgctcc tcctaatctc cgagctcttc ccgggctcca tccctatgga gccccatctg
     2761 gcatggctct cccctataac cccatgatgg tagttatgat gcctccaccc ccaccccctg
     2821 tctccacagc agtgcagccc cctggtgccc ctccagtcag agacctgggc tccgtgcccc
     2881 cagaactgac agctagccgt cagagcttcc acatggccat gggcaacccc agtgagtttt
     2941 ttgtggatgt tatgtagctc gagccatgga agcttgatat ctaactgact gaaccggtgg
     3001 taccaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3061 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3121 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3181 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3241 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3301 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3361 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3421 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3481 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3541 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3601 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3661 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3721 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4081 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     4141 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca
     4201 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     4261 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     4321 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     4381 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     4441 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     4501 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     4561 ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4621 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4681 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4861 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7081 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7141 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7201 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     7261 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
