LOCUS       sequence-252105-        5838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of mouse Spop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5838)
  AUTHORS   Cai H, Liu A
  TITLE     Spop promotes skeletal development and homeostasis by positively 
            regulating Ihh signaling.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Dec 20;113(51):14751-14756. doi: 
            10.1073/pnas.1612520114. Epub 2016 Dec 5.
  PUBMED    27930311
REFERENCE   2  (bases 1 to 5838)
  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"; doi: 
            "10.1073/pnas.1612520114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-12-20- 20"; volume: "113"; issue: "51"; pages: 
            "14751-14756"
COMMENT     Sequence Label: pEGFPC1-Spop
FEATURES             Location/Qualifiers
     source          1..5838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2262..2383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2299..2318)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2353..2372
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2390..2845)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2527..2548)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2739..2758
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2872..2976
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2978..3335
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2978..3169
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3001..3021)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3186..3321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3370..4164
                     /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             3370..4164
                     /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(3424..3443)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4034..4053
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4352..4371)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4396..4443
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4772..5360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5261..5280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5535..5838
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgag tcaagggttc
     1021 caagtcctcc acctccggca gaaatgtcga gtggtcccgt tgccgagagc tggtgctata
     1081 cacagatcaa ggtagtgaag ttctcctaca tgtggaccat caataacttt agtttttgcc
     1141 gggaggaaat gggtgaagtc ataaaaagtt caactttttc atcaggagcc aatgataaac
     1201 tgaaatggtg tttgcgagta aacccgaaag ggctagatga agaaagcaaa gattacctgt
     1261 cgctttacct gttgttggtc agctgtccaa agagtgaagt tcgggcaaag ttcaagttct
     1321 ccatcctgaa tgccaaggga gaagaaacca aagctatgga gagtcagcga gcttataggt
     1381 tcgtgcaagg caaagactgg ggattcaaaa aattcatccg cagagatttt ctcttggacg
     1441 aggccaatgg gcttctccct gatgacaagc tcaccctctt ctgtgaggtg agtgtagtac
     1501 aagactctgt caatatttct gggcagaata ctatgaacat ggtaaaggtt cctgaatgcc
     1561 ggttggcaga tgagttagga gggctgtggg agaactctcg attcaccgac tgctgcttgt
     1621 gtgtggctgg tcaagaattc caggctcaca aggctatctt agcagctcgg tctccagtct
     1681 tcagtgccat gtttgaacat gaaatggagg agagcaaaaa gaatcgggtt gagattaacg
     1741 acgtggagcc tgaggtgttt aaggagatga tgtgcttcat ctacacgggg aaggcgccga
     1801 acctcgacaa gatggctgac gacttgctgg cagctgccga taagtacgcc ctggagcgtc
     1861 tgaaggtcat gtgtgaggac gccctctgca gtaacctctc tgtggagaac gctgcagaga
     1921 ttctcatcct ggctgacctc cacagcgcag atcagctgaa gacgcaggca gttgatttca
     1981 tcaactatca tgcttcggat gtcttggaga cctctgggtg gaagtcaatg gtggtgtcac
     2041 atccccactt agtggcagaa gcataccgtt ctctggcttc agcacagtgc cctttcctgg
     2101 ggcccccacg caaacgcctg aagcaatcct aaggatccac cggatctaga taactgatca
     2161 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2221 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2281 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2341 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     2401 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2461 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2521 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2581 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2641 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2701 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2761 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2821 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2881 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2941 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3001 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3061 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3121 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3181 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3241 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3301 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3361 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3421 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3481 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3541 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3601 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3661 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3721 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3781 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3841 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3901 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3961 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4021 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4081 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4141 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4201 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4261 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4321 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4381 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4441 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4501 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4561 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4621 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4681 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4741 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4801 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4861 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4921 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4981 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5041 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5101 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5161 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5221 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5281 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5341 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5401 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5461 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     5521 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     5581 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     5641 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5701 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5761 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     5821 gtcatcgcta ttaccatg
//
