LOCUS       sequence-238728-        6417 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 6417)
  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 6417)
  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-Ubxd8
FEATURES             Location/Qualifiers
     source          1..6417
                     /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      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      970..979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2373..2390)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2379..2603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2649..3077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2736..2755)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2946..2967
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3086..3106)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3091..3420
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3271..3406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3333..3352
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3468..3515
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3534..3932
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4090..4211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4127..4146)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4181..4200
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4260..4276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4260..4276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4273..4295)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4284..4300
                     /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(4308..4314,4315..4332,4333..4338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4353..4374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4491..4508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4662..5250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4742..4761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5421..6212,6213..6281))
                     /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     6044..6063
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6282..6386)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagt
      901 tagagctcct taaggttaac gccaccatgg actacaaaga cgatgacgac aagtctagag
      961 aattcggatc catggcggcg cctgaggagc aggatctaac tcaggagcag accgagaagc
     1021 tgctgcagtt tcaggatctg actggcatag agtccatgga gcagtgtcgc cttgctttgg
     1081 agcagcataa ctggaacatg gaggctgctg tccaggacag actgaatgag caggaaggtg
     1141 taccaagtgt tttcaatcca ccgccagctc ggcccctaca ggttaataca gctgaccaca
     1201 ggatctacag ctatgttgtc tcaagaccac aaccaagggg cctgcttgga tggggttatt
     1261 acttgataat gcttccattc cggtttacct attacacgat acttgatata tttaggtttg
     1321 ctcttcgttt tatacggcct gaccctcgca gccgggtcac tgaccctgtt ggggacattg
     1381 tttcatttat gcactctttt gaagagaaat atgggagggc acaccctgtc ttctaccagg
     1441 gaacgtacag ccaggcactt aacgatgcca agcgggaact tcgatttctt ttggtttatc
     1501 ttcatggaga tgaccaccag gactctgatg agttctgccg caatgctctc tgcgcacctg
     1561 aggtcatctc actgataaac agcaggatgc ttttctgggc ttgttctaca aacaagcctg
     1621 aggggtacag ggtgtcccag gccttacggg agaacaccta tccattccta gctatgatta
     1681 tgttgaagga tcggcgaatg actgtggttg gaaggctaga aggcctcatt caacctgatg
     1741 acctcatcaa ccagctgaca ttcatcatgg atgcaaacca gacttacctg gtgtcagaac
     1801 gtctagaaag ggaagaaagg aaccaaaccc aggtgctgag acaacagcag gatgaagcct
     1861 acctggcttc tctcagggca gatcaggaga aagagaggaa gaagagggag gagaaagagc
     1921 ggaaacgccg gaaggaggag gaggtgcagc aacagaagct ggcggaggag cgaagacgac
     1981 agaacttgca ggaggaaaag gagaggaagc tggaatgcct gccccctgag ccctcccctg
     2041 atgatcctga aagtgtcaag atcattttca agttacccaa tgattccaga gtagagagac
     2101 gattccactt ctcacagtct ctaacagtca tccatgactt cttattctcc ttgaaggaaa
     2161 gccctgaaaa gtttcagatt gaagccaatt ttccaagacg ggtacttccc tgtgttcctt
     2221 cagaggaatg gcccaacccc ccaacactgc aggaggcggg actcagccac acagaagttc
     2281 tctttgtcca ggacctgaca gacgaataac tcgagccatg gaagcttgat atctaactga
     2341 ctgaaccggt ggtacctaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2401 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2461 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2521 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2581 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2641 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2701 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2761 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2821 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2881 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2941 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3001 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3061 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3241 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3301 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3361 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3421 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     3481 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     3541 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     3601 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     3661 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     3721 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     3781 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     3841 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     3901 tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga
     3961 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4021 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4081 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4141 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4201 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4261 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4321 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4381 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4441 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4501 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4561 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4621 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4681 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4741 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4801 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4861 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4921 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4981 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5041 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5101 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5161 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5221 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5281 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5341 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5401 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5461 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5521 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5581 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5641 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5701 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5761 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5821 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5881 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5941 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6001 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6061 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6121 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6181 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6241 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6301 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6361 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
