LOCUS       sequence-244133-        7577 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 7577)
  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 7577)
  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-Axin1
FEATURES             Location/Qualifiers
     source          1..7577
                     /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(3299..3316)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3305..3529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3575..4003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3662..3681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3872..3893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4012..4032)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4017..4346
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4197..4332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4259..4278
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4394..4441
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4460..4858
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5016..5137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5053..5072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5107..5126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5186..5202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5186..5202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5199..5221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5210..5226
                     /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(5234..5240,5241..5258,5259..5264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5279..5300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5417..5434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5588..6176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5668..5687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6347..7138,7139..7207))
                     /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     6970..6989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7208..7312)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7387..7406)
                     /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 cagagtccca aaatgaatgt ccaggagcag ggtttcccct
      781 tggacctcgg agcaagtttc accgaagatg ccccccggcc cccagtgcct ggagaagagg
      841 gagaactggt atctactgat tcgaggcctg tcaaccacag tttctgttct gggaaaggta
      901 ccagcattaa aagtgagacc tcaacagcca ccccaagacg ttcagatctg gatctgggat
      961 atgagcccga gggcagtgcc tcccccaccc caccatattt gaggtgggct gagtcactgc
     1021 attccttact ggatgaccaa gatgggatca gcctgttcag gactttcctg aagcaggagg
     1081 gctgtgctga cctgctggac ttctggtttg cctgcagtgg cttcaggaag cttgagccct
     1141 gtgactcaaa tgaggaaaag aggctgaagc tggcaagagc catctaccga aagtacatcc
     1201 tggatagcaa tggcattgtg tccagacaaa ccaagccagc cactaagagc ttcataaagg
     1261 actgtgtcat gaagcagcag atagatcctg ccatgtttga ccaggcacag acagaaatcc
     1321 agtccaccat ggaggagaat acctaccctt cctttcttaa gtctgacatt tatttggagt
     1381 acacaaggac aggctcagag agtccgaagg tctgcagtga ccagagctca gggtctggaa
     1441 cagggaaggg catgtctgga tacctgccca ctttgaatga ggatgaagaa tggaaatgtg
     1501 accaagatgc agatgaggat gatggccgag accctctccc ccccagcagg ctcacccaga
     1561 agctgctatt ggagactgct gccccgaggg ccccctcaag tagacggtac aacgaaggca
     1621 gagagctcag gtatggatct tggagggagc ccgtcaaccc ctactacgtc aactctggct
     1681 atgccctggc cccagccacc agtgccaatg acagtgagca gcagagcctg tccagtgatg
     1741 ctgacacgct atcccttacg gacagtagtg tggatggaat ccccccatac aggatccgta
     1801 agcagcaccg aagggagatg caggagagta tccaagtcaa tgggcgggta cctctacctc
     1861 acattcctcg cacttaccga atgccaaagg agatccgggt agagccacag aaatttgctg
     1921 aagagcttat tcaccgtcta gaggctgtcc agcgcactcg agaggctgaa gaaaagttgg
     1981 aggaacggct gaagcgtgta cgcatggagg aagaagggga ggatggtgaa atgccttctg
     2041 gccccatggc aagtcacaag ctgccttctg tcccagcttg gcaccatttc ccaccccgct
     2101 atgtggatat gggctgctct ggactgcggg atgcccatga ggagaatcct gagagcatcc
     2161 tggatgagca cgtgcaaagg gtcatgagga cacctggctg ccagtcacct ggcccaggcc
     2221 accgctctcc tgacagtggg catgtggcta agactgcagt gctagggggt acagcctccg
     2281 ggcatgggaa gcatgttcct aagttagggt tgaagctgga tacagctggc ctgcaccatc
     2341 atagacatgt ccaccaccat gttcaccata attcagctag acctaaggag caaatggagg
     2401 ctgaagttgc ccgcagggtc cagagcagct tctcgtgggg cccagaaaca catggtcatg
     2461 ccaagccccg gagctattcc gagaacgcag gcaccaccct cagtgctggg gatttggcct
     2521 ttggtggtaa aactagtgca ccttccaaaa gaaacaccaa gaaggctgaa tctgggaaga
     2581 acgccaatgc tgaggtaccc agtaccacag aggacgctga gaagaaccag aagatcatgc
     2641 agtggatcat tgagggagag aaggagatca gtagacaccg gaaggcaggc catgggtctt
     2701 ctgggttgag gaagcagcag gcccatgaaa gctccaggcc cttgtccatc gagcgtcctg
     2761 gggccgtgca cccctgggtc agcgctcagc ttcggaattc tgtccagcct tctcatcttt
     2821 tcatccaaga tcccacaatg ccacccaatc cagcccctaa tcccctgacc cagctggaag
     2881 aggcccgcag gcgtttggaa gaagaagaaa agagagcaaa caaactgccc tccaagcaga
     2941 ggacaaaatc acaaagaaag gcaggtggcg ggagtgcacc accatgtgac agcattgttg
     3001 tggcctacta tttctgtggg gaacccatcc cctaccggac cctggtgagg ggccgtgctg
     3061 tcaccctggg ccagttcaag gagctgctaa ccaagaaggg gagctacaga tactacttta
     3121 agaaagtgag tgatgagttt gactgtggtg tggtatttga ggaagtacgg gaggatgagg
     3181 ccgtcttgcc tgtctttgaa gaaaagatca tcggcaaggt ggaaaaggtg gactgactcg
     3241 agccatggaa gcttgatatc taactgactg aaccggtggt accaaacccg ctgatcagcc
     3301 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3361 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3421 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3481 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3541 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3601 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3661 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3721 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3781 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3841 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3901 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3961 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4261 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4321 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4381 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     4441 caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     4501 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     4561 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     4621 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     4681 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     4741 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     4801 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc
     4861 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     4921 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4981 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5161 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5221 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5281 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5341 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5401 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5461 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5521 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5581 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5641 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5701 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5761 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5821 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5881 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5941 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6001 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6061 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6121 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6181 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6241 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6301 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6361 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6421 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6481 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6541 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6601 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6661 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6721 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6781 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6841 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6901 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6961 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7021 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7081 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7141 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7201 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7261 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7321 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     7381 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     7441 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     7501 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     7561 ggccagatat acgcgtt
//
