LOCUS       sequence-251106-        7424 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 7424)
  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 7424)
  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-Ctnnb1
FEATURES             Location/Qualifiers
     source          1..7424
                     /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"
     CDS             783..818
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     primer_bind     complement(3146..3163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3152..3376
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3422..3850
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3509..3528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3719..3740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3859..3879)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3864..4193
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4044..4179
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4106..4125
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4241..4288
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4307..4705
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4863..4984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4900..4919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4954..4973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5033..5049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5033..5049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5046..5068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5057..5073
                     /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(5081..5087,5088..5105,5106..5111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5126..5147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5264..5281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5435..6023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5515..5534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6194..6985,6986..7054))
                     /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     6817..6836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7055..7159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7234..7253)
                     /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 cggatccatg gctactcaag ctgacctgat ggagttggac atggccatgg
      781 agccggacag aaaagctgct gtcagccact ggcagcagca gtcttacttg gattctggaa
      841 tccattctgg tgccaccacc acagctcctt ccctgagtgg caagggcaac cctgaggaag
      901 aagatgttga cacctcccaa gtcctttatg aatgggagca aggcttttcc cagtccttca
      961 cgcaagagca agtagctgat attgacgggc agtatgcaat gactagggct cagagggtcc
     1021 gagctgccat gttccctgag acgctagatg agggcatgca gatcccatcc acgcagtttg
     1081 acgctgctca tcccactaat gtccagcgct tggctgaacc atcacagatg ttgaaacatg
     1141 cagttgtcaa tttgattaac tatcaggatg acgcggaact tgccacacgt gcaattcctg
     1201 agctgacaaa actgctaaac gatgaggacc aggtggtagt taataaagct gctgttatgg
     1261 tccatcagct ttccaaaaag gaagcttcca gacatgccat catgcgctcc cctcagatgg
     1321 tgtctgccat tgtacgcacc atgcagaata caaatgatgt agagacagct cgttgtactg
     1381 ctgggactct gcacaacctt tctcaccacc gcgagggctt gctggccatc tttaagtctg
     1441 gtggcatccc agcgctggtg aaaatgcttg ggtcaccagt ggattctgta ctgttctacg
     1501 ccatcacgac actgcataat ctcctgctcc atcaggaagg agctaaaatg gcagtgcgcc
     1561 tagctggtgg actgcagaaa atggttgctt tgctcaacaa aacaaacgtg aaattcttgg
     1621 ctattacaac agactgcctt cagatcttag cttatggcaa tcaagagagc aagctcatca
     1681 ttctggccag tggtggaccc caagccttag taaacataat gaggacctac acttatgaga
     1741 agcttctgtg gaccacaagc agagtgctga aggtgctgtc tgtctgctct agcaacaagc
     1801 cggccattgt agaagctggt gggatgcagg cactggggct tcatctgaca gacccaagtc
     1861 agcgacttgt tcaaaactgt ctttggactc tcagaaacct ttcagatgca gcgactaagc
     1921 aggaagggat ggaaggcctc cttgggactc tagtgcagct tctgggttcc gatgatataa
     1981 atgtggtcac ctgtgcagct ggaattctct ctaacctcac ttgcaataat tacaaaaaca
     2041 agatgatggt gtgccaagtg ggtggcatag aggctcttgt acgcaccgtc cttcgtgctg
     2101 gtgacaggga agacatcact gagcctgcca tctgtgctct tcgtcatctg accagccggc
     2161 atcaggaagc cgagatggcc cagaatgccg ttcgccttca ttatggactg cctgttgtgg
     2221 ttaaactcct gcacccacca tcccactggc ctctgataaa ggcaactgtt ggattgattc
     2281 gaaaccttgc cctttgccca gcaaatcatg cgcctttgcg ggaacagggt gctattccac
     2341 gactagttca gctgcttgta cgagcacatc aggacaccca acggcgcacc tccatgggtg
     2401 gaacgcagca gcagtttgtg gagggcgtgc gcatggagga gatagtagaa gggtgtactg
     2461 gagctctcca catccttgct cgggacgttc acaaccggat tgtaatccga ggactcaata
     2521 ccattccatt gtttgtgcag ttgctttatt ctcccattga aaatatccaa agagtagctg
     2581 caggggtcct ctgtgaactt gctcaggaca aggaggctgc agaggccatt gaagctgagg
     2641 gagccacagc tcccctgaca gagttactcc actccaggaa tgaaggcgtg gcaacatacg
     2701 cagctgctgt cctattccga atgtctgagg acaagccaca ggattacaag aagcggcttt
     2761 cagtcgagct gaccagttcc ctcttcagga cagagccaat ggcttggaat gagactgcag
     2821 atcttggact ggacattggt gcccagggag aagcccttgg atatcgccag gatgatccca
     2881 gctaccgttc ttttcactct ggtggatacg gccaggatgc cttggggatg gaccctatga
     2941 tggagcatga gatgggtggc caccaccctg gtgctgacta tccagttgat gggctgcctg
     3001 atctgggaca cgcccaggac ctcatggatg ggctgccccc aggtgatagc aatcagctgg
     3061 cctggtttga tactgacctg taactcgagc catggaagct tgatatctaa ctgactgaac
     3121 cggtggtacc aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3181 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3241 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3301 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3361 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3421 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3481 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3541 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3601 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3661 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3721 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3781 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3841 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4021 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4081 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4141 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4201 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4261 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt
     4321 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     4381 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     4441 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     4501 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     4561 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     4621 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     4681 ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc
     4741 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4801 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4861 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4921 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4981 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5041 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5101 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6241 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6601 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     7201 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7261 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     7321 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     7381 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
