LOCUS       sequence-244173-        7208 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 7208)
  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 7208)
  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-Fzd6
FEATURES             Location/Qualifiers
     source          1..7208
                     /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"
     primer_bind     complement(2930..2947)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2936..3160
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3206..3634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3503..3524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3643..3663)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3648..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..4489
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4647..4768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4684..4703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4738..4757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4817..4833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4817..4833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4830..4852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4841..4857
                     /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(4865..4871,4872..4889,4890..4895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4910..4931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5048..5065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5219..5807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5299..5318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5978..6769,6770..6838))
                     /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     6601..6620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6839..6943)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7018..7037)
                     /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 gaaaggtccc cgtttctgtt ggcgtgcatt cttctgcccc
      781 tcgtaagagg acacagcctt ttcacctgtg agccaatcac cgttcccaga tgtatgaaaa
      841 tgacttacaa catgacgttc ttccctaacc tgatgggtca ttatgaccag gggatcgctg
      901 ctgtggaaat ggggcacttt ctgcatcttg caaatctaga atgttcacca aacattgaaa
      961 tgttcctttg ccaagctttt ataccaacct gcacagagca aattcatgta gttctaccct
     1021 gtcggaaatt gtgtgagaaa atagtttctg attgcaaaaa actaatggac acttttggca
     1081 tccgatggcc tgaagaactt gaatgtaaca gattgccaca ctgtgatgac actgttcctg
     1141 taacttctca tccacacaca gagctttctg ggccacagaa gaaatcagat caagtcccaa
     1201 gagacattgg attttggtgt ccaaagcacc ttaggacttc cggggaccaa ggctataggt
     1261 ttctgggaat tgaacagtgt gcccctccgt gccccaatat gtattttaaa agtgatgaac
     1321 tagactttgc caaaagtttc ataggaatag tttcaatatt ttgtctttgt gcaactctgt
     1381 tcacgttcct tacattttta attgacgtta gacgattcag atacccagag agaccaatta
     1441 tctattactc tgtctgctac agcattgtct ctctcatgta cttcgtgggg tttttgctgg
     1501 gcaatagcac ggcttgtaat aaggcagacg agaagctgga gctcggggac accgttgtcc
     1561 tagggtcaaa gaataaggct tgcagtgtgg tatttatgtt tctgtatttt tttacaatgg
     1621 ctggcaccgt gtggtgggtg attctcacca ttacgtggtt cttagctgcc gggagaaaat
     1681 ggagttgcga agctattgaa caaaaagcag tgtggttcca tgccgttgcc tggggggcgc
     1741 ccgggttcct gaccgtcatg ctgctcgcta tgaataaggt tgaaggagac aacattagcg
     1801 gcgtttgctt cgttggcctg tatgacctgg acgcctctcg ctacttcgtc cttctgcctc
     1861 tgtgcctctg cgtatttgtt gggctgtctc tcctcttagc cggcatcatc tccttgaatc
     1921 atgtccgaca agtcatacag catgatggcc ggaaccaaga gaagctaaag aaattcatga
     1981 ttcgcatcgg agtcttcagt ggcctgtatc ttgtgccctt agtgacactt ctcggttgct
     2041 atgtctatga gctagtgaac aggatcacct gggagatgac atggttctct gatcattgtc
     2101 accagtaccg catcccgtgc ccttaccagg caaatccaaa agctcgacca gaattggctt
     2161 tatttatgat aaaatatctg atgacattaa ttgttggtat ctctgcggtc ttctgggttg
     2221 gaagcaaaaa gacgtgcaca gaatgggccg ggttctttaa gcgaaaccgc aagcgagacc
     2281 ccatcagtga gagccgccga gtgctgcaag agtcctgtga gttcttcctg aagcacaact
     2341 ctaaagtgaa gcacaagaag aagcatggcg caccagggcc tcataggctg aaggtcattt
     2401 ccaagtccat gggaactagc acaggagcga ccacaaatca tggcacctct gccatggcaa
     2461 tcgctgacca tgattactta gggcaagaaa cttcaacaga agtccacacc tccccagaag
     2521 catccgtcaa agagggacga gcagaccgag caaacactcc cagcgccaaa gatcgggact
     2581 gtggggaatc tgcagggccc agttccaagc tctctgggaa ccggaacggc agggaaagcc
     2641 gagcgggcgg cctgaaggag agaagcaatg gatcagaggg ggctccaagt gaaggaaggg
     2701 taagtccaaa gagcagcgtt cctgagactg gcctgataga ctgcagcact tcacaggccg
     2761 ccagttctcc agaaccaacc agcctcaagg gctccacatc tctgcctgtt cactcagctt
     2821 ccagagctag gaaagagcag ggtgctggca gccattccga cgcttgactc gagccatgga
     2881 agcttgatat ctaactgact gaaccggtgg taccaaaccc gctgatcagc ctcgactgtg
     2941 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3001 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3061 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3121 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3181 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3661 ggtgtggaaa gtccccaggc tccccagcag 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 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     4141 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     4201 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     4261 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     4321 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     4381 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     4441 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     4501 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4561 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4621 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4681 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4741 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4801 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4861 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4921 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4981 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5041 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5101 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6121 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6841 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6961 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     7021 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     7081 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     7141 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     7201 tacgcgtt
//
