LOCUS       sequence-255696-        6998 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 6998)
  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 6998)
  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-Gp78
FEATURES             Location/Qualifiers
     source          1..6998
                     /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(2720..2737)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2726..2950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2996..3424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3083..3102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3293..3314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3433..3453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3438..3767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3618..3753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3680..3699
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3815..3862
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3881..4279
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4437..4558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4474..4493)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4528..4547
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4607..4623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4607..4623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4620..4642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4631..4647
                     /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(4655..4661,4662..4679,4680..4685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4700..4721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4838..4855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5009..5597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5089..5108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5768..6559,6560..6628))
                     /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     6391..6410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6629..6733)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6808..6827)
                     /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 ccgctgctct tcctcgagcg cttcccgtgg cccagcctcc
      781 gcacctacac gggcctcagc ggcctggcct tgctgggcac catcgtcagc gcctaccgcg
      841 ccctcagtca gccggaggac ggctcgggtg agccggagcc gctaacggcc ccactgcagc
      901 ccgaggcgcg cctgaccgcc ggcggccccc gggcccgcga cgtggcccag tacctgctgt
      961 cggacagcct gttcgtgtgg gttctggtga atactgcttg ctgtgttttg atgttggtgg
     1021 ctaagcttat ccagtgtatt gtgttcggcc ctcttcgagt gagtgaaaga cagcatctca
     1081 aagacaagtt ttggaatttt attttctaca agttcatttt catttttgga gtgctgaatg
     1141 tccagactgt ggaggaagtg gtgatgtggt gcctctggtt tgctggactt gtcttcctgc
     1201 atctgatggt tcagctctgc aaagacagat ttgaatacct ttccttttct cccaccactc
     1261 caatgagcag ccatgggaga gtcctgtccc tgctgattgc tatgctgctc tcctgctgtg
     1321 gactggcagt cgtctgctgt gtcacaggct acacacatgg catgcacacc ttggctttca
     1381 tggctgcaga gtctcttctt gtgaccgtga gaacagctca tgtgatttta agatatgtaa
     1441 ttcacctctg ggacctcaac cacgaaggga cgtgggaagg aaagggaacc tatgtgtatt
     1501 acaccgactt tgtcatggag ctcgctctcc tgtccctgga cctcatgcac cacatccaca
     1561 tgctgttatt tggcaacatc tggttatcta tggctagctt ggtcatcttt atgcaacttc
     1621 gttacctgtt ccatgaggta cagcgtcgaa tccgtcgaca caagaattat ttgcgagtgg
     1681 taggaaacat ggaagccagg tttgctgttg caactccaga ggaactagct gtcaataatg
     1741 atgactgtgc aatctgctgg gactccatgc aggctgcaag gaaactgccc tgtggacatc
     1801 tttttcacaa ctcctgcctt cgttcttggc tagaacaaga cacctcttgt ccaacatgca
     1861 gaatgtctct taatattgct gatggcagtc gtgccaggga agaccatcaa ggagagaacc
     1921 tggacgagaa cttggttcct gtagcagctg ccgaaggcag gcctcgcttg aaccagcaca
     1981 atcacttctt ccacttcgat gggtcccgga ttgcaagctg gctgccaagt ttttcagttg
     2041 aagtgatgca cactaccaac attcttggca tcacgcaggc aagcaactct cagctaaatg
     2101 ccatggctca tcagattcag gagatgttcc ctcaggtccc gtaccacctt gtgctccagg
     2161 acctccagat gacacgctcc gtggagataa caacagataa catcttagaa ggacggattc
     2221 aagtaccttt ccccacgcag cggtcggata gcctgaggcc tgcactgaac agcccagtgg
     2281 agaggcccag ccctgacctg gaggagggag aagcttctgt ccagacggag cgtgtaccac
     2341 tggacctcag tcctcgccta gaggagacct tggacttcag tgaggtagaa ctagagccta
     2401 ttgaggtgga agactttgag gcccgtggaa gccgcttctc caagtctgct gatgagagac
     2461 agcgcatgtt agttcagcgc aaggatgacc tcctgcagca ggctcgaaag cggttcttaa
     2521 acaaaagttc tgaagatgat ggggcctcag agagactcct cccttctgaa ggcacatcct
     2581 ctgaccccgt gaccctgcgc cgaaggatgc tggcagccgc tgctgagcgg agacttcaga
     2641 ggcaacggac aacctagctc gagccatgga agcttgatat ctaactgact gaaccggtgg
     2701 taccaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2761 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2821 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2881 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2941 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3001 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3061 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3181 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3301 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3361 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3421 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3781 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     3841 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca
     3901 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     3961 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     4021 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     4081 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     4141 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     4201 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     4261 ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4321 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4381 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4441 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4501 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4561 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4621 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4681 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4741 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4801 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4861 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4921 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4981 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5041 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5101 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5161 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5221 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5281 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5341 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5401 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5461 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5521 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6721 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6781 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6841 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6901 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6961 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
