LOCUS       sequence-244134-        6341 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 6341)
  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 6341)
  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-Gsk3b
FEATURES             Location/Qualifiers
     source          1..6341
                     /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     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(2063..2080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2069..2293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2339..2767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2426..2445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2636..2657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2776..2796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2781..3110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2961..3096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3023..3042
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3158..3205
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3224..3622
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3780..3901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3817..3836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3871..3890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3950..3966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3950..3966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3963..3985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3974..3990
                     /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(3998..4004,4005..4022,4023..4028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4043..4064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4181..4198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4432..4451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5111..5902,5903..5971))
                     /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     5734..5753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5972..6076)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6151..6170)
                     /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 tcggggcgac cgagaaccac ctcctttgcg gagagctgca
      781 agccagtgca gcagccttca gcttttggta gcatgaaagt tagcagagat aaagatggca
      841 gcaaggtaac cacagtagtg gcaactcctg gccagggtcc tgacaggcca caggaagtca
      901 gttatacaga cacgaaagtg attggaaatg gatcatttgg tgtggtatat caagccaaac
      961 tttgtgattc tggagaactg gttgccatca agaaagttct acaggacaag cgatttaaga
     1021 accgagagct ccagatcatg agaaagctag accactgtaa catagtccga ctgcggtatt
     1081 tcttctactc gagtggtgag aagaaagatg aggtctacct taacctggtg ctggactatg
     1141 ttccggagac agtgtacaga gtcgccagac actatagtcg agccaagcag acactccctg
     1201 tgatctatgt caagttgtat atgtatcagc tgttcagaag tctagcctat atccattcct
     1261 ttggaatctg ccatcgagac attaaaccac agaacctctt gttggatcct gatacagctg
     1321 tattaaaact ctgtgacttt ggaagtgcaa agcagctggt ccgaggagag cccaatgttt
     1381 catatatctg ttctcggtac tacagggcac cagagttgat ctttggagcc actgattaca
     1441 cgtccagtat agatgtatgg tctgcaggct gtgtgttggc tgaattgttg ctaggacaac
     1501 caatatttcc tggggacagt ggtgtggatc agttggtgga aataataaag gtcctaggaa
     1561 caccaacaag ggagcaaatt agagaaatga acccaaatta tacagaattc aaattccctc
     1621 aaatcaaggc acatccttgg acaaaggtct tccggccccg aactccacca gaggcaattg
     1681 cactgtgcag ccgtctgctg gagtacacac ctaccgcccg gctaacacca ctggaagctt
     1741 gtgcacattc atttttcgat gaattgcggg acccaaatgt caaactacca aatgggcgag
     1801 acacacctgc actcttcaac tttaccactc aagaactgtc gagtaacccc cctctggcca
     1861 ccatccttat ccctccacat gctcggattc aggccgctgc ttcaccgcct gccaacgcca
     1921 cagcagcctc agatactaat gctggagacc gtggacagac caataacgcc gcttctgcat
     1981 cagcttccaa ctccacctga ctcgagccat ggaagcttga tatctaactg actgaaccgg
     2041 tggtaccaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2101 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2161 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2221 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2281 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2341 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2401 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2461 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2521 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2581 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2641 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2701 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2761 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2941 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3001 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3061 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3121 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     3181 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc
     3241 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     3301 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     3361 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     3421 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     3481 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     3541 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     3601 tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga cacgtgctac
     3661 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3721 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3901 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3961 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4021 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4081 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4141 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5101 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5161 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5221 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5281 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5341 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5401 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5461 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5521 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5581 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5641 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5701 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5761 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5821 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5881 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5941 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6001 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6061 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6121 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6181 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     6241 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     6301 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
