LOCUS       sequence-244132-        7499 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 7499)
  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 7499)
  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-Vcp
FEATURES             Location/Qualifiers
     source          1..7499
                     /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(3221..3238)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3227..3451
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3497..3925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3584..3603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3794..3815
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3934..3954)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3939..4268
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4119..4254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4181..4200
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4316..4363
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4382..4780
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4938..5059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4975..4994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5029..5048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5108..5124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5108..5124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5121..5143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5132..5148
                     /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(5156..5162,5163..5180,5181..5186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5201..5222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5339..5356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5510..6098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5590..5609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6269..7060,7061..7129))
                     /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     6892..6911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7130..7234)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7309..7328)
                     /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 gcctctggag ccgattcaaa aggtgatgat ttatcaacag
      781 ccattctcaa acagaagaac cgacccaatc ggttaattgt tgatgaagcc atcaatgaag
      841 ataacagcgt ggtgtccttg tcccagccca agatggatga actgcagttg ttccgaggtg
      901 acacggtgtt gctaaaagga aagaaaagac gggaagctgt atgcattgtt ctttctgatg
      961 acacgtgttc tgatgagaag attcgaatga atagagttgt tcggaataac ctccgagttc
     1021 gcctaggaga tgtcatcagc atccagccat gccctgatgt aaagtatggc aaacgtatcc
     1081 acgttctacc catcgatgac acagtggaag gcatcactgg caatctcttt gaggtatacc
     1141 ttaagccgta cttcctggaa gcttatcggc ccatccgtaa aggagatatt tttcttgtcc
     1201 ggggtgggat gcgtgctgtg gagttcaaag ttgtagagac agatcccagc ccttactgta
     1261 ttgttgctcc agacacagtg atccactgtg agggggagcc aatcaagcga gaggatgagg
     1321 aggaatcctt gaatgaagta ggctatgatg acatcggtgg ttgcaggaag cagctagctc
     1381 agataaagga gatggtggag ctgccactga gacatcctgc gctctttaag gcgattggtg
     1441 taaagcctcc tcggggaatc ttgttgtatg ggcctcctgg gacagggaag accctgattg
     1501 ctcgagctgt ggcaaatgaa actggagcct tcttctttct gatcaatggt cctgaaatca
     1561 tgagcaaatt ggctggtgag tctgagagca accttcgtaa agcctttgag gaagctgaaa
     1621 agaatgctcc tgctatcatc ttcatcgatg agcttgatgc cattgcaccc aaaagagaga
     1681 aaactcatgg ggaagtggag cgtcgcatcg tgtctcagtt gttgaccctc atggatggcc
     1741 taaagcagag agcacatgtg atagttatgg cagcaaccaa tagacccaac agcattgacc
     1801 cagccctacg gcgatttggt cgctttgaca gagaggtaga tattggaata cctgatgcta
     1861 caggacgttt ggagattctt cagatccata ccaagaacat gaaactggca gatgatgtgg
     1921 acttggaaca ggtagccaat gagactcatg gtcatgttgg tgctgatttg gcagccctat
     1981 gttcagaggc tgctctgcag gccatccgga aaaaaatgga cctcattgac ctagaagatg
     2041 agaccattga tgctgaggtc atgaattccc tggcagttac tatggatgac ttccggtggg
     2101 ctttgagtca aagcaaccca tcagcacttc gggaaactgt ggtagaggtg ccacaagtaa
     2161 cctgggaaga tattggaggc ctggaggatg tcaaacgtga gcttcaggag ttggttcagt
     2221 atcctgtgga acatccagac aaattcctca aatttggcat gactccctcc aaaggcgttc
     2281 ttttctatgg acctcctggc tgtgggaaaa ccttactggc taaagccatt gctaatgaat
     2341 gccaggccaa cttcatctcc atcaagggtc ctgagctgct taccatgtgg tttggggaat
     2401 ctgaggccaa tgtccgggaa atttttgaca aggcacggca agctgccccc tgtgtactct
     2461 tctttgatga gttagattca attgccaagg ctcgaggtgg taatattgga gatggtggtg
     2521 gagctgctga ccgagtcatc aatcagatcc tgacagaaat ggatggcatg tctacaaaaa
     2581 agaatgtgtt tatcattgga gctaccaaca ggcctgacat cattgatcct gctatcctaa
     2641 gacctggccg tctagatcag ctcatttata tcccacttcc tgatgagaag tcccgtgttg
     2701 ccatcctaaa agccaatctg cgaaagtccc cagttgccaa ggatgtggat ttggagttcc
     2761 tggctaagat gactaatggc ttttctggag ctgatttgac agaaatttgc caacgggctt
     2821 gtaaactggc cattcgtgaa tctattgaga gtgagattag gcgagaacga gagaggcaga
     2881 caaatccatc ggctatggag gtagaagagg acgatccagt gcctgagatc cgcagagatc
     2941 actttgagga agccatgcgt tttgcccgac gttctgtcag cgataatgac attcggaagt
     3001 atgaaatgtt cgcccagaca ctgcagcaga gtcgaggttt tggcagcttc agattccctt
     3061 cagggaacca gggtggagct ggtcccagcc agggcagtgg aggtggcaca ggtggcagtg
     3121 tgtacacaga agacaatgac gatgacctgt atggctaact cgagccatgg aagcttgata
     3181 tctaactgac tgaaccggtg gtaccaaacc cgctgatcag cctcgactgt gccttctagt
     3241 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3301 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3361 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3421 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3481 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3781 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3841 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3901 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4141 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4201 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4261 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     4321 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     4381 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     4441 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     4501 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     4561 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     4621 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     4681 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     4741 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc
     4801 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4921 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4981 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5041 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7261 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     7321 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7381 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     7441 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
