LOCUS       sequence-362439-        7880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Flag tagged expression clone for the mouse Prom1 splicing 
            variant SV8 (GenBank accession BC028286). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7880)
  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 7880)
  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
FEATURES             Location/Qualifiers
     source          1..7880
                     /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"
     CDS             3425..3448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3471..3488)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3477..3701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3747..4175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3834..3853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4044..4065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4184..4204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4189..4518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4369..4504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4431..4450
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4585..5379
                     /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(4639..4658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5249..5268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5553..5674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5590..5609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5644..5663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5723..5739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5723..5739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5736..5758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5747..5763
                     /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(5771..5777,5778..5795,5796..5801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5816..5837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5954..5971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6125..6713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6205..6224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6884..7675,7676..7744))
                     /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     7507..7526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7745..7849)
                     /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 gctccaccat
      901 ggctctcgtc ttcagtgccc tgctgttact ggggctgtgt ggaaagatct cttcagaagg
      961 tcagcctgca ttccataaca ctcctggggc tatgaattat gaattgccta ccaccaaata
     1021 tgagacccaa gataccttca atgctgggat tgttggccct ctctacaaaa tggtgcacat
     1081 cttcctcaac gtggtccagc cgaatgactt ccctctagat ttgatcaaaa aactcataca
     1141 gaacaagaac tttgacatct cagttgattc caaggagcca gaaatcatag tcttggctct
     1201 gaagattgcc ctctatgaga tcggagtcct tatctgcgcc atcctgggac tgctgttcat
     1261 tatcctcatg cctctggtgg gctgcttctt ttgtatgtgc cgttgctgca acaaatgcgg
     1321 cggagagatg caccagcggc agaagcagaa tgcgccatgc aggaggaagt gcttgggcct
     1381 ctccctcctg gtgatttgtc tgctcatgag ccttggcatt atatatggct ttgtggctaa
     1441 ccagcagacc aggactcgga tcaaagggac ccagaaactg gcaaagagca atttcagaga
     1501 ctttcaaaca ctcctgactg aaacaccaaa gcaaattgac tatgtagtgg agcagtacac
     1561 caacaccaag aacaaggcat tctcagacct ggatggcatc ggctccgtgc tgggaggcag
     1621 aataaaggac caactaaaac ccaaagtaac tcctgtcctc gaagagatta aggccatggc
     1681 gacagccatc aaacagacca aggatgccct gcagaacatg agcagcagcc tgaaaagtct
     1741 ccaagatgca gccacccagc tcaataccaa cctgagctct gtgagaaaca gcatcgagaa
     1801 ttcgctcagc agcagtgact gtacctcaga tccagccagc aagatctgcg atagcatcag
     1861 accaagccta agcagtctgg ggagcagcct caattcaagt cagctcccat cagtggatag
     1921 agaactcaac actgttactg aagtcgacaa aactgatctg gagagcctcg tcaaaagggg
     1981 gtatacgaca attgatgaaa tacccaatac aatacaaaac caaactgtgg atgtcatcaa
     2041 agacgtcaaa aataccttgg actccattag ctccaacatt aaggacatga gccaaagtat
     2101 tcctattgag gatatgctgt tacaggtctc ccattacctt aataacagca acagatactt
     2161 aaaccaggag ctgcccaagc tggaagaata tgactcgtac tggtggctgg gtggcttgat
     2221 tgtctgcttt ctgctgactc tcattgtgac cttctttttc ctgggcttgc tgtgtggtgt
     2281 gtttggctat gacaagcatg ccaccccaac tagaagaggc tgtgtgtcca acactggagg
     2341 catcttcctc atggctgggg ttggattcgg cttccttttt tgctggatat tgatgatcct
     2401 tgtggttctt acgtttgttg ttggtgcaaa tgtggaaaag ttgctctgcg aaccttatga
     2461 aaacaagaaa ttattacagg ttttggacac tccctatctg ctcaaggaac aatggcaatt
     2521 ttatctttct ggcatgctat tcaataaccc agacattaac atgacctttg agcaagtcta
     2581 cagggattgc aaaagaggtc gaggtatata tgctgctttt cagcttgaga atgtcgtcaa
     2641 cgtcagtgat catttcaaca ttgaccagat ttctgaaaac ataaatacgg agttggaaaa
     2701 cctgaatgtg aacattgata gcattgaact gttggataac acaggaagga agagcctcga
     2761 ggactttgca cattctggga tagatacaat cgattattcc acatacttga aggagactga
     2821 gaaatcccct actgaagtga atctgctgac atttgcctct accctggaag caaaagcaaa
     2881 ccagttgcct gaaggaaagc tgaaacaggc cttcttactg gatgtacaga atataagagc
     2941 catccaccag catctcctcc ctcctgtgca gcaatcactg aaatttgtga gggtgaggaa
     3001 tacgttaaga caaagtgtct ggaccctcca gcaaacaagc aacaagttgc cggagaaagt
     3061 gaagaagatc cttgcctctt tggactctgt tcagcatttc ctcaccaata acgtttccct
     3121 catcgttatc ggggaaacga agaagtttgg gaaaacaata ctaggctact ttgaacatta
     3181 tctgcactgg gtcttttatg ccatcacaga gaagatgaca tcctgcaaac ccatggccac
     3241 cgcgatggac tctgctgtta atggcattct gtgtggctat gttgcggacc ctctgaattt
     3301 gttctggttc ggcataggga aagccacggt gctcttactt ccggctgtaa tcattgctat
     3361 caagctggcc aagtactatc gcaggatgga ttcagaggat gtatacgacg acccgtctcg
     3421 atacgactac aaagacgatg acgacaagtg agtttaaacc cgctgatcag cctcgactgt
     3481 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3541 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3601 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3661 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3721 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3781 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3841 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3901 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3961 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4021 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4081 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4141 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4201 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4261 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4321 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4381 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4441 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4501 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4561 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4621 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4681 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4741 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4801 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4861 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4921 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4981 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5041 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5101 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5161 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5221 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5281 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5341 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5401 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5461 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5521 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5641 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5701 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5761 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5821 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5881 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5941 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6001 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6061 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7021 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7861 gaaaagtgcc acctgacgtc
//
