LOCUS       sequence-244135-        6539 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 6539)
  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 6539)
  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-C-3XFLAG-h-Lmbr1l
FEATURES             Location/Qualifiers
     source          1..6539
                     /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"
     RBS             1716..1724
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2193..2216
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2261..2278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2267..2491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2537..2965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2624..2643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2834..2855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2974..2994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2979..3308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3159..3294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3221..3240
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3356..3403
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3422..3820
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3978..4099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4015..4034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4069..4088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4148..4164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4148..4164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4161..4183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4172..4188
                     /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(4196..4202,4203..4220,4221..4226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4241..4262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4379..4396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4550..5138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4630..4649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5309..6100,6101..6169))
                     /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     5932..5951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6170..6274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6349..6368)
                     /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 gttaactcta gagaattcgg atccatggaa gcacctgact
      721 acgaagtgct atccgtgcga gaacagctat tccacgagag gatccgcgag tgtattatat
      781 caacacttct gtttgcaaca ctgtacatcc tctgccacat cttcctgacc cgcttcaaga
      841 agcctgctga gttcaccaca gtggatgatg aagatgccac cgtcaacaag attgcgctcg
      901 agctgtgcac ctttaccctg gcaattgccc tgggtgctgt cctgctcctg cccttctcca
      961 tcatcagcaa tgaggtgctg ctctccctgc ctcggaacta ctacatccag tggctcaacg
     1021 gctccctcat ccatggcctc tggaaccttg tttttctctt ctccaacctg tccctcatct
     1081 tcctcatgcc ctttgcatat ttcttcactg agtctgaggg ctttgctggc tccagaaagg
     1141 gtgtcctggg ccgggtctat gagacagtgg tgatgttgat gctcctcact ctgctggtgc
     1201 taggtatggt gtgggtggca tcagccattg tggacaagaa caaggccaac agagagtcac
     1261 tctatgactt ttgggagtac tatctcccct acctctactc atgcatctcc ttccttgggg
     1321 ttctgctgct cctggtgtgt actccactgg gtctcgcccg catgttctcc gtcactggga
     1381 agctgctagt caagccccgg ctgctggaag acctggagga gcagctgtac tgctcagcct
     1441 ttgaggaggc agccctgacc cgcaggatct gtaatcctac ttcctgctgg ctgcctttag
     1501 acatggagct gctacacaga caggtcctgg ctctgcagac acagagggtc ctgctggaga
     1561 agaggcggaa ggcttcagcc tggcaacgga acctgggcta ccccctggct atgctgtgct
     1621 tgctggtgct gacgggcctg tctgtgctca ttgtggccat ccacatcctg gagctgctca
     1681 tcgatgaggc tgccatgccc cgaggcatgc agggtacctc cttaggccag gtctccttct
     1741 ccaagctggg ctcctttggt gccgtcattc aggttgtact catcttttac ctaatggtgt
     1801 cctcagttgt gggcttctat agctctccac tcttccggag cctgcggccc agatggcacg
     1861 acactgccat gacgcagata attgggaact gtgtctgtct cctggtccta agctcagcac
     1921 ttcctgtctt ctctcgaacc ctggggctca ctcgctttga cctgctgggt gactttggac
     1981 gcttcaactg gctgggcaat ttctacattg tgttcctcta caacgcagcc tttgcaggcc
     2041 tcaccacact ctgtctggtg aagaccttca ctgcagctgt gcgggcagag ctgatccggg
     2101 cctttgggct ggacagactg ccgctgcccg tctccggttt cccccaggca tctaggaaga
     2161 cccagcacca gctcgagcca tggaagctta tggactacaa agacgatgac gacaaggata
     2221 tctaactgac tgaaccggtg gtaccaaacc cgctgatcag cctcgactgt gccttctagt
     2281 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2341 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2401 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2461 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2521 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2581 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2641 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2701 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2761 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2821 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2881 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2941 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3181 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3241 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3301 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     3361 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3421 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     3481 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     3541 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     3601 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     3661 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     3721 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     3781 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc
     3841 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3901 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3961 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4081 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6301 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     6361 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6421 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6481 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
