LOCUS       sequence-244129-        6119 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 6119)
  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 6119)
  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-HA-Ubac2
FEATURES             Location/Qualifiers
     source          1..6119
                     /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"
     CDS             696..722
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1841..1858)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1847..2071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2117..2545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2204..2223)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2414..2435
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2554..2574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2559..2888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2739..2874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2801..2820
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2936..2983
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3002..3400
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3558..3679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3595..3614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3649..3668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3728..3744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3728..3744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3741..3763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3752..3768
                     /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(3776..3782,3783..3800,3801..3806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3821..3842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3959..3976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4130..4718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4210..4229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4889..5680,5681..5749))
                     /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     5512..5531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5750..5854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5929..5948)
                     /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 ccatgtaccc atacgatgtt ccagattacg
      721 cttctagaga attcggatcc atgttcacca gcaccggctc cagtgggctt tataaggcac
      781 ctctgtcgaa gagtcttctg ctggtcccca gtgccctgtc cctcctgctg actctcctgc
      841 tgcctcactg tcagaagttc ttcgtgtatg acctccatgc tgtcaagcac gacttgcaga
      901 tttggaggct gatatgcgga agaataattt gccttgactt gaaagacgct ttctgtagtg
      961 gtcttctcat ttataacttc agaatatttg aaagaagata tggaagcagg aagttcgcat
     1021 ccttcttgct gggttcctgg gtcttgtcag ccttgtttga ttttatcctc gtggaagccg
     1081 tgcagtattc gcttggtgtc acagtggcca gcaacttacc ttctggattc ttggcgcccg
     1141 tgttcgctct ctttgttcca tttcactgct ccatcccaag agtccaggtg gcgcagatcc
     1201 tgggcccgtt gtccatcaca aacaagacat tgatttatat attgggacta cagcttttca
     1261 cctctgggtc ctacatctgg attgtagcca tgagtggact tatctctggc atgtgctatg
     1321 accgcaaagt gctccaggtc caccaggtgc tccgcatccc tggcagaatg gccgagttct
     1381 tctcctgggc ccttgagccc atcttctcat cttcagaacc caccagcgag gccagggttg
     1441 ggatgggggc caccgtggac atccagagac agcagaggat ggagcagctg gaccggcagc
     1501 tgatgctctc tcagtttgca caagtgaggc gtcaaaggca gcagcaggga ggaatgatca
     1561 attggaaccg gcttttcccc cctttacgtc agcgacgaaa tatcaactat caagatggac
     1621 ctcggtctga gcagcgagca tctcctcccc tggaagtttc tgaagagcag gttgcccgcc
     1681 tcatggagat gggattttcc agaggggacg ccttggaggc cctgagggcc tcaaacaatg
     1741 acctcaatgt ggccaccaac ttcctgctgc agcactgact cgagccatgg aagcttgata
     1801 tctaactgac tgaaccggtg gtaccaaacc cgctgatcag cctcgactgt gccttctagt
     1861 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1921 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1981 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2041 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2101 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2161 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2221 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2281 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2341 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2401 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2461 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2521 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2581 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2641 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2761 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2821 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2881 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     2941 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3001 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     3061 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     3121 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     3181 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     3241 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     3301 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     3361 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc
     3421 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3481 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3541 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3601 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3661 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     3721 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3781 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3841 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3901 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3961 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4021 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4081 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4141 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4201 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4261 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4321 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4381 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4441 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4501 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4561 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4621 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4681 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4741 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4801 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4861 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4921 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4981 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5041 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5101 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5161 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5221 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5281 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5341 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5401 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5461 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5521 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5581 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5641 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5701 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5761 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5821 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5881 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     5941 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6001 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6061 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
