LOCUS       sequence-244131-        6416 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 6416)
  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 6416)
  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..6416
                     /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"
     regulatory      736..745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2138..2155)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2144..2368
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2414..2842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2501..2520)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2711..2732
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2851..2871)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2856..3185
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3036..3171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3098..3117
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3233..3280
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3299..3697
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3855..3976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3892..3911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3946..3965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4025..4041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4025..4041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4038..4060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4049..4065
                     /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(4073..4079,4080..4097,4098..4103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4118..4139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4256..4273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4427..5015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4507..4526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5186..5977,5978..6046))
                     /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     5809..5828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6047..6151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6226..6245)
                     /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 gcggcgcctg aggagcagga tctaactcag gagcagaccg
      781 agaagctgct gcagtttcag gatctgactg gcatagagtc catggagcag tgtcgccttg
      841 ctttggagca gcataactgg aacatggagg ctgctgtcca ggacagactg aatgagcagg
      901 aaggtgtacc aagtgttttc aatccaccgc cagctcggcc cctacaggtt aatacagctg
      961 accacaggat ctacagctat gttgtctcaa gaccacaacc aaggggcctg cttggatggg
     1021 gttattactt gataatgctt ccattccggt ttacctatta cacgatactt gatatattta
     1081 ggtttgctct tcgttttata cggcctgacc ctcgcagccg ggtcactgac cctgttgggg
     1141 acattgtttc atttatgcac tcttttgaag agaaatatgg gagggcacac cctgtcttct
     1201 accagggaac gtacagccag gcacttaacg atgccaagcg ggaacttcga tttcttttgg
     1261 tttatcttca tggagatgac caccaggact ctgatgagtt ctgccgcaat gctctctgcg
     1321 cacctgaggt catctcactg ataaacagca ggatgctttt ctgggcttgt tctacaaaca
     1381 agcctgaggg gtacagggtg tcccaggcct tacgggagaa cacctatcca ttcctagcta
     1441 tgattatgtt gaaggatcgg cgaatgactg tggttggaag gctagaaggc ctcattcaac
     1501 ctgatgacct catcaaccag ctgacattca tcatggatgc aaaccagact tacctggtgt
     1561 cagaacgtct agaaagggaa gaaaggaacc aaacccaggt gctgagacaa cagcaggatg
     1621 aagcctacct ggcttctctc agggcagatc aggagaaaga gaggaagaag agggaggaga
     1681 aagagcggaa acgccggaag gaggaggagg tgcagcaaca gaagctggcg gaggagcgaa
     1741 gacgacagaa cttgcaggag gaaaaggaga ggaagctgga atgcctgccc cctgagccct
     1801 cccctgatga tcctgaaagt gtcaagatca ttttcaagtt acccaatgat tccagagtag
     1861 agagacgatt ccacttctca cagtctctaa cagtcatcca tgacttctta ttctccttga
     1921 aggaaagccc tgaaaagttt cagattgaag ccaattttcc aagacgggta cttccctgtg
     1981 ttccttcaga ggaatggccc aaccccccaa cactgcagga ggcgggactc agccacacag
     2041 aagttctctt tgtccaggac ctgacagacg aatgactcga gccatggaag cttgatatct
     2101 aactgactga accggtggta ccaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2161 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2221 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2281 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2341 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2401 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2461 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2521 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2581 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2641 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2701 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2761 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2821 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2881 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2941 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3001 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3061 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3121 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3181 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     3241 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     3301 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     3361 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     3421 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     3481 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     3541 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     3601 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     3661 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg
     3721 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3781 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3841 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3901 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3961 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4021 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4081 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4141 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4201 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4261 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4321 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4381 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4441 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4501 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4561 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4621 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4681 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4741 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4801 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4861 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4921 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4981 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5041 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5101 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5161 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5221 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5281 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5341 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5401 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5461 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5521 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5581 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5641 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5701 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5761 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5821 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5881 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5941 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6001 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6061 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6121 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6181 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     6241 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     6301 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6361 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
