LOCUS       sequence-362442-        7863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Flag tagged expression clone for the mouse Prom1 splicing 
            variant SV8 (GenBank accession BC028286). Deletion of exon 19a 
            (amino acids 696-701)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7863)
  AUTHORS   Matalkah F, Rhodes S, Ramamurthy V, Stoilov P
  TITLE     The mAB 13A4 monoclonal antibody to the mouse PROM1 protein 
            recognizes a structural epitope.
  JOURNAL   PLoS One. 2022 Oct 10;17(10):e0274958. doi: 
            10.1371/journal.pone.0274958. eCollection 2022.
  PUBMED    36215230
REFERENCE   2  (bases 1 to 7863)
  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: "PLoS One.";
            date: "2022-10-10"; volume: "17(10):e0274958. doi"; pages: " 
            10.1371/journal.pone.0274958. eCollection 2022"
COMMENT     Sequence Label: pcDNA-Prom1 SV8 (-Ex19a)
FEATURES             Location/Qualifiers
     source          1..7863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3408..3431
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3460..3684
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3730..4158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3817..3836)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4027..4048
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4167..4187)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4172..4501
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4352..4487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4414..4433
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4568..5362
                     /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(4622..4641)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5232..5251
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5536..5657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5573..5592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5627..5646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5706..5722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5706..5722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5719..5741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5730..5746
                     /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(5754..5760,5761..5778,5779..5784))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5799..5820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5937..5954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6108..6696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6188..6207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6867..7658,7659..7727))
                     /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     7490..7509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7728..7832)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctgccacca
      901 tggctctcgt cttcagtgcc ctgctgttac tggggctgtg tggaaagatc tcttcagaag
      961 gtcagcctgc attccataac actcctgggg ctatgaatta tgaattgcct accaccaaat
     1021 atgagaccca agataccttc aatgctggga ttgttggccc tctctacaaa atggtgcaca
     1081 tcttcctcaa cgtggtccag ccgaatgact tccctctaga tttgatcaaa aaactcatac
     1141 agaacaagaa ctttgacatc tcagttgatt ccaaggagcc agaaatcata gtcttggctc
     1201 tgaagattgc cctctatgag atcggagtcc ttatctgcgc catcctggga ctgctgttca
     1261 ttatcctcat gcctctggtg ggctgcttct tttgtatgtg ccgttgctgc aacaaatgcg
     1321 gcggagagat gcaccagcgg cagaagcaga atgcgccatg caggaggaag tgcttgggcc
     1381 tctccctcct ggtgatttgt ctgctcatga gccttggcat tatatatggc tttgtggcta
     1441 accagcagac caggactcgg atcaaaggga cccagaaact ggcaaagagc aatttcagag
     1501 actttcaaac actcctgact gaaacaccaa agcaaattga ctatgtagtg gagcagtaca
     1561 ccaacaccaa gaacaaggca ttctcagacc tggatggcat cggctccgtg ctgggaggca
     1621 gaataaagga ccaactaaaa cccaaagtaa ctcctgtcct cgaagagatt aaggccatgg
     1681 cgacagccat caaacagacc aaggatgccc tgcagaacat gagcagcagc ctgaaaagtc
     1741 tccaagatgc agccacccag ctcaatacca acctgagctc tgtgagaaac agcatcgaga
     1801 attcgctcag cagcagtgac tgtacctcag atccagccag caagatctgc gatagcatca
     1861 gaccaagcct aagcagtctg gggagcagcc tcaattcaag tcagctccca tcagtggata
     1921 gagaactcaa cactgttact gaagtcgaca aaactgatct ggagagcctc gtcaaaaggg
     1981 ggtatacgac aattgatgaa atacccaata caatacaaaa ccaaactgtg gatgtcatca
     2041 aagacgtcaa aaataccttg gactccatta gctccaacat taaggacatg agccaaagta
     2101 ttcctattga ggatatgctg ttacaggtct cccattacct taataacagc aacagatact
     2161 taaaccagga gctgcccaag ctggaagaat atgactcgta ctggtggctg ggtggcttga
     2221 ttgtctgctt tctgctgact ctcattgtga ccttcttttt cctgggcttg ctgtgtggtg
     2281 tgtttggcta tgacaagcat gccaccccaa ctagaagagg ctgtgtgtcc aacactggag
     2341 gcatcttcct catggctggg gttggattcg gcttcctttt ttgctggata ttgatgatcc
     2401 ttgtggttct tacgtttgtt gttggtgcaa atgtggaaaa gttgctctgc gaaccttatg
     2461 aaaacaagaa attattacag gttttggaca ctccctatct gctcaaggaa caatggcaat
     2521 tttatctttc tggcatgcta ttcaataacc cagacattaa catgaccttt gagcaagtct
     2581 acagggattg caaaagaggt cgaggtatat atgctgcttt tcagcttgag aatgtcgtca
     2641 acgtcagtga tcatttcaac attgaccaga tttctgaaaa cataaatacg gagttggaaa
     2701 acctgaatgt gaacattgat agcattgaac tgttggataa cacaggaagg aagagcctcg
     2761 aggactttgc acattctggg atagatacaa tcgattattc cacatacttg aaggagactg
     2821 agaaatcccc tactgaagtg aatctgctga catttgcctc taccctggaa gcaaaagcaa
     2881 accagttgcc tgaaggaaag ctgaaacagg ccttcttact ggatgtacag aatataagag
     2941 ccatccacca gcatctcctc cctcctgtgc agcaatcact gaatacgtta agacaaagtg
     3001 tctggaccct ccagcaaaca agcaacaagt tgccggagaa agtgaagaag atccttgcct
     3061 ctttggactc tgttcagcat ttcctcacca ataacgtttc cctcatcgtt atcggggaaa
     3121 cgaagaagtt tgggaaaaca atactaggct actttgaaca ttatctgcac tgggtctttt
     3181 atgccatcac agagaagatg acatcctgca aacccatggc caccgcgatg gactctgctg
     3241 ttaatggcat tctgtgtggc tatgttgcgg accctctgaa tttgttctgg ttcggcatag
     3301 ggaaagccac ggtgctctta cttccggctg taatcattgc tatcaagctg gccaagtact
     3361 atcgcaggat ggattcagag gatgtatacg acgacccgtc tcgatacgac tacaaagacg
     3421 atgacgacaa gtgagtttaa acccgctgat cagcatcgac tgtgccttct agttgccagc
     3481 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3541 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3601 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3661 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3721 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3781 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3841 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3901 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3961 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4021 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4081 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4141 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4321 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4381 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4441 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4501 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4561 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4741 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5341 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5401 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5461 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5521 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5581 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5641 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7861 gtc
//
