LOCUS       sequence-498401-        7303 bp    DNA     circular SYN 28-JAN-2026
DEFINITION  Express mouse sp7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7303)
  AUTHORS   Wu L, Wang L, Zhuang Y, Luo Y, Zhu Q, Maimaitizunong M, Wang Y, 
            Cheng Z, Li Y, Sheng X, Li M, Luo Q, Jiang X, Lei S, Lin X, Zhong Y,
            Ren W
  TITLE     Integrative analysis reveals synergistic regulation of Sp7 by BRD9 
            and Wnt/beta-catenin signaling during osteogenic differentiation.
  JOURNAL   Commun Biol. 2025 Dec 1. doi: 10.1038/s42003-025-09278-z.
  PUBMED    41326797
REFERENCE   2  (bases 1 to 7303)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 28, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol. 2025 Dec 1. doi: 10.1038/s42003-025-09278-z."
COMMENT     Sequence Label: pCMV-sp7(mouse)-3HA
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      677..686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1979..2005
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2009..2035
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2039..2065
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2081..2669
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2134..2154)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2552..2563)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2691..2710)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2735..2783
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2985..3413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3072..3091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3282..3303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3422..3442)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3427..3756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3607..3742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3669..3688
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3823..4617
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3877..3896)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4487..4506
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4791..4912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4828..4847)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4882..4901
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4961..4977)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4961..4977)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4974..4996)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4985..5001
                     /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(5009..5015,5016..5033,5034..5039))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5054..5075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5192..5209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5363..5951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5443..5462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6122..6913,6914..6982))
                     /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     6745..6764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6983..7087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7162..7181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcgtc ctctctgctt gaggaagaag ctcactatgg
      721 ctccagtccc ctggccatgc tgactgcagc ctgcagcaaa tttggcggct ctagccctct
      781 gcgggactca acaaccctgg ggaaaggagg cacaaagaag ccatacgctg acctttcagc
      841 ccccaaaacc atgggggacg cctacccagc tcccttctca agcaccaatg gactcctctc
      901 tcctgcaggc agtcctccgg ccccagcctc tggctatgca aatgactacc cacccttccc
      961 tcactcattt cctgggccca ccggtgccca agaccctggg ctcctagtgc ctaaggggca
     1021 cagctcgtct gactgcctgc ctagtgtcta cacttccctg gatatgactc atccctatgg
     1081 ctcgtggtac aaggcaggca tccacgcagg catctcacca ggtccaggca acacacctac
     1141 tccttggtgg gacatgcacc ctgggggcaa ctggctaggt ggtggtcagg gccagggtga
     1201 tgggctgcaa gggacactgt ccacaggccc tgcccagcct ccactgaacc cccagctgcc
     1261 tacttaccca tctgactttg ctccccttaa cccagctccc tacccagcgc cccacctctt
     1321 gcaaccaggg ccccagcatg tcctacccca agatgtctat aagcccaagg cggttggcaa
     1381 tagtgggcaa ctggagggga gtggtgcagc caaaccccct cggggtgctg gcacaggggg
     1441 cagcggtgga tatgcgggca gtggggcagg gcgttctacc tgcgactgcc ccaactgtca
     1501 ggagctagag cggctcgggg cagcagcggc tgggctgagg aagaagccca ttcacagctg
     1561 ccacatccct gggtgcggca aggtgtacgg caaggcttcg catctgaaag cccacttgcg
     1621 ctggcacact ggcgagaggc ctttcgtctg caactggctt ttctgcggca agaggttcac
     1681 tcgctctgac gagctggagc gccacgtgcg cactcacacc cgggagaaga agttcacctg
     1741 cctgctctgt tccaagcgct ttaccagaag cgaccacttg agcaaacatc agcgcaccca
     1801 cggggagcca ggcccgggac cgcccccaag tggccctaag gagctggggg agggtcgcag
     1861 cgtcggggaa gaagaagcca atcagccgcc ccgatcttcc acttcgcctg cacccccaga
     1921 aaaagcccac ggaggcagcc cagagcagag caacctgcta gagatcctcg agggtaccta
     1981 cccatacgac gtacctgatt atgcaactta tccttatgat gtacctgatt atgctagcta
     2041 tccttatgat gtcccagact acgcatagtg atatctcgac aatcaacctc tggattacaa
     2101 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2161 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2221 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2281 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2341 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2401 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2461 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     2521 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2581 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2641 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     2701 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     2761 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     2821 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     2881 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     2941 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3421 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3661 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3781 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4321 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4621 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4681 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4801 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4861 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4921 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4981 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5041 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5101 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5521 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5761 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5881 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5941 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6001 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6061 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6121 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6181 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6241 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6301 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6361 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6421 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6481 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6541 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6601 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6661 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6721 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6781 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6841 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6901 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6961 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7021 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7081 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7141 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     7201 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     7261 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
