LOCUS       sequence-244126-        6821 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 6821)
  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 6821)
  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: pEGFP-N1-C-Fzd6
FEATURES             Location/Qualifiers
     source          1..6821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2701..2710
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2707..2709,2710..2712,2713..3426)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2707..2709,2710..2712,2713..3423)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3013..3032)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3360..3381
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3549..3670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3586..3605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3640..3659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3677..4132)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3814..3835)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4026..4045
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4159..4263
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4265..4622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4265..4456
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4288..4308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4473..4608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4657..5451
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4657..5451
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4711..4730)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5321..5340
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5639..5658)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5683..5730
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6059..6647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6548..6567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagat ggaaaggtcc ccgtttctgt tggcgtgcat tcttctgccc
      601 ctcgtaagag gacacagcct tttcacctgt gagccaatca ccgttcccag atgtatgaaa
      661 atgacttaca acatgacgtt cttccctaac ctgatgggtc attatgacca ggggatcgct
      721 gctgtggaaa tggggcactt tctgcatctt gcaaatctag aatgttcacc aaacattgaa
      781 atgttccttt gccaagcttt tataccaacc tgcacagagc aaattcatgt agttctaccc
      841 tgtcggaaat tgtgtgagaa aatagtttct gattgcaaaa aactaatgga cacttttggc
      901 atccgatggc ctgaagaact tgaatgtaac agattgccac actgtgatga cactgttcct
      961 gtaacttctc atccacacac agagctttct gggccacaga agaaatcaga tcaagtccca
     1021 agagacattg gattttggtg tccaaagcac cttaggactt ccggggacca aggctatagg
     1081 tttctgggaa ttgaacagtg tgcccctccg tgccccaata tgtattttaa aagtgatgaa
     1141 ctagactttg ccaaaagttt cataggaata gtttcaatat tttgtctttg tgcaactctg
     1201 ttcacgttcc ttacattttt aattgacgtt agacgattca gatacccaga gagaccaatt
     1261 atctattact ctgtctgcta cagcattgtc tctctcatgt acttcgtggg gtttttgctg
     1321 ggcaatagca cggcttgtaa taaggcagac gagaagctgg agctcgggga caccgttgtc
     1381 ctagggtcaa agaataaggc ttgcagtgtg gtatttatgt ttctgtattt ttttacaatg
     1441 gctggcaccg tgtggtgggt gattctcacc attacgtggt tcttagctgc cgggagaaaa
     1501 tggagttgcg aagctattga acaaaaagca gtgtggttcc atgccgttgc ctggggggcg
     1561 cccgggttcc tgaccgtcat gctgctcgct atgaataagg ttgaaggaga caacattagc
     1621 ggcgtttgct tcgttggcct gtatgacctg gacgcctctc gctacttcgt ccttctgcct
     1681 ctgtgcctct gcgtatttgt tgggctgtct ctcctcttag ccggcatcat ctccttgaat
     1741 catgtccgac aagtcataca gcatgatggc cggaaccaag agaagctaaa gaaattcatg
     1801 attcgcatcg gagtcttcag tggcctgtat cttgtgccct tagtgacact tctcggttgc
     1861 tatgtctatg agctagtgaa caggatcacc tgggagatga catggttctc tgatcattgt
     1921 caccagtacc gcatcccgtg cccttaccag gcaaatccaa aagctcgacc agaattggct
     1981 ttatttatga taaaatatct gatgacatta attgttggta tctctgcggt cttctgggtt
     2041 ggaagcaaaa agacgtgcac agaatgggcc gggttcttta agcgaaaccg caagcgagac
     2101 cccatcagtg agagccgccg agtgctgcaa gagtcctgtg agttcttcct gaagcacaac
     2161 tctaaagtga agcacaagaa gaagcatggc gcaccagggc ctcataggct gaaggtcatt
     2221 tccaagtcca tgggaactag cacaggagcg accacaaatc atggcacctc tgccatggca
     2281 atcgctgacc atgattactt agggcaagaa acttcaacag aagtccacac ctccccagaa
     2341 gcatccgtca aagagggacg agcagaccga gcaaacactc ccagcgccaa agatcgggac
     2401 tgtggggaat ctgcagggcc cagttccaag ctctctggga accggaacgg cagggaaagc
     2461 cgagcgggcg gcctgaagga gagaagcaat ggatcagagg gggctccaag tgaaggaagg
     2521 gtaagtccaa agagcagcgt tcctgagact ggcctgatag actgcagcac ttcacaggcc
     2581 gccagttctc cagaaccaac cagcctcaag ggctccacat ctctgcctgt tcactcagct
     2641 tccagagcta ggaaagagca gggtgctggc agccattccg acgctatgga tccaccggtc
     2701 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2761 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2821 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2881 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2941 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3001 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3061 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3121 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3181 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3241 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3301 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3361 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3421 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3481 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3541 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3601 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3661 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3721 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3781 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3841 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3901 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3961 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4021 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4081 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4141 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4201 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4261 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4381 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4441 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4501 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4561 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4621 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4681 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4741 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4801 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4861 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4921 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4981 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5041 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5101 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5161 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5221 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5281 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5341 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5401 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5461 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5521 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5581 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5641 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5701 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5761 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5821 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5881 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5941 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6001 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6121 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6181 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6541 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6601 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6661 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6721 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
     6781 attacggggt cattagttca tagcccatat atggagttcc g
//
