LOCUS       sequence-362420-        7830 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), and deletion of amino acids 661 to 675..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7830)
  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 7830)
  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 D-3
FEATURES             Location/Qualifiers
     source          1..7830
                     /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      896..905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3362..3385
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3421..3438)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3427..3651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3697..4125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3784..3803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3994..4015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4134..4154)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4139..4468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4319..4454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4381..4400
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4535..5329
                     /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(4589..4608)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5199..5218
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5503..5624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5540..5559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5594..5613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5673..5689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5673..5689)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5686..5708)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5697..5713
                     /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(5721..5727,5728..5745,5746..5751))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5766..5787
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5904..5921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6075..6663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6155..6174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6834..7625,7626..7694))
                     /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     7457..7476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7695..7799)
                     /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 gctgggccac
      901 catggctctc gtcttcagtg ccctgctgtt actggggctg tgtggaaaga tctcttcaga
      961 aggtcagcct gcattccata acactcctgg ggctatgaat tatgaattgc ctaccaccaa
     1021 atatgagacc caagatacct tcaatgctgg gattgttggc cctctctaca aaatggtgca
     1081 catcttcctc aacgtggtcc agccgaatga cttccctcta gatttgatca aaaaactcat
     1141 acagaacaag aactttgaca tctcagttga ttccaaggag ccagaaatca tagtcttggc
     1201 tctgaagatt gccctctatg agatcggagt ccttatctgc gccatcctgg gactgctgtt
     1261 cattatcctc atgcctctgg tgggctgctt cttttgtatg tgccgttgct gcaacaaatg
     1321 cggcggagag atgcaccagc ggcagaagca gaatgcgcca tgcaggagga agtgcttggg
     1381 cctctccctc ctggtgattt gtctgctcat gagccttggc attatatatg gctttgtggc
     1441 taaccagcag accaggactc ggatcaaagg gacccagaaa ctggcaaaga gcaatttcag
     1501 agactttcaa acactcctga ctgaaacacc aaagcaaatt gactatgtag tggagcagta
     1561 caccaacacc aagaacaagg cattctcaga cctggatggc atcggctccg tgctgggagg
     1621 cagaataaag gaccaactaa aacccaaagt aactcctgtc ctcgaagaga ttaaggccat
     1681 ggcgacagcc atcaaacaga ccaaggatgc cctgcagaac atgagcagca gcctgaaaag
     1741 tctccaagat gcagccaccc agctcaatac caacctgagc tctgtgagaa acagcatcga
     1801 gaattcgctc agcagcagtg actgtacctc agatccagcc agcaagatct gcgatagcat
     1861 cagaccaagc ctaagcagtc tggggagcag cctcaattca agtcagctcc catcagtgga
     1921 tagagaactc aacactgtta ctgaagtcga caaaactgat ctggagagcc tcgtcaaaag
     1981 ggggtatacg acaattgatg aaatacccaa tacaatacaa aaccaaactg tggatgtcat
     2041 caaagacgtc aaaaatacct tggactccat tagctccaac attaaggaca tgagccaaag
     2101 tattcctatt gaggatatgc tgttacaggt ctcccattac cttaataaca gcaacagata
     2161 cttaaaccag gagctgccca agctggaaga atatgactcg tactggtggc tgggtggctt
     2221 gattgtctgc tttctgctga ctctcattgt gaccttcttt ttcctgggct tgctgtgtgg
     2281 tgtgtttggc tatgacaagc atgccacccc aactagaaga ggctgtgtgt ccaacactgg
     2341 aggcatcttc ctcatggctg gggttggatt cggcttcctt ttttgctgga tattgatgat
     2401 ccttgtggtt cttacgtttg ttgttggtgc aaatgtggaa aagttgctct gcgaacctta
     2461 tgaaaacaag aaattattac aggttttgga cactccctat ctgctcaagg aacaatggca
     2521 attttatctt tctggcatgc tattcaataa cccagacatt aacatgacct ttgagcaagt
     2581 ctacagggat tgcaaaagag gtcgaggtat atatgctgct tttcagcttg agaatgtcgt
     2641 caacgtcagt gatcatttca acattgacca gatttctgaa aacataaata cggagttgga
     2701 aaacctgaat gtgaacattg atagcattga actgttggat aacacaggaa ggaagagcct
     2761 cgaggacttt gcacattctg ggatagatac aatcgattat tccacatact tgaaggagac
     2821 tgagaaatcc cctactgaag tgaatctgct gacatttgcc tctaccctgg aagcaaaagt
     2881 acagaatata agagccatcc accagcatct cctccctcct gtgcagcaat cactgaatac
     2941 gttaagacaa agtgtctgga ccctccagca aacaagcaac aagttgccgg agaaagtgaa
     3001 gaagatcctt gcctctttgg actctgttca gcatttcctc accaataacg tttccctcat
     3061 cgttatcggg gaaacgaaga agtttgggaa aacaatacta ggctactttg aacattatct
     3121 gcactgggtc ttttatgcca tcacagagaa gatgacatcc tgcaaaccca tggccaccgc
     3181 gatggactct gctgttaatg gcattctgtg tggctatgtt gcggaccctc tgaatttgtt
     3241 ctggttcggc atagggaaag ccacggtgct cttacttccg gctgtaatca ttgctatcaa
     3301 gctggccaag tactatcgca ggatggattc agaggatgta tacgacgacc cgtctcgata
     3361 cgactacaaa gacgatgacg acaagtgagt ctagagggcc cgtttaaacc cgctgatcag
     3421 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3481 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3541 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3601 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3661 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3721 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3781 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3841 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3901 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3961 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4021 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4081 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4141 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4201 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4261 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4321 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4381 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4441 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4501 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4561 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4621 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4681 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4741 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4801 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4861 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4921 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4981 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5041 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5101 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5161 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5221 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5281 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5341 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5401 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5461 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5521 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5581 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5641 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5701 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5761 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5821 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5881 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5941 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6001 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6061 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6121 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6181 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6241 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6301 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6361 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6421 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6481 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6541 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6601 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6661 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6721 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6781 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6841 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6901 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6961 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7021 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7081 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7141 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7201 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7261 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7321 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7381 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7441 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7501 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7561 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7621 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7681 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7741 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7801 acatttcccc gaaaagtgcc acctgacgtc
//
