LOCUS       sequence-362422-        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 amino acids 
            685-706..
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-1
FEATURES             Location/Qualifiers
     source          1..7830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..48)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        33..362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      213..348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     275..294
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             429..1223
                     /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(483..502)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1093..1112
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1397..1518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1434..1453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1488..1507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1567..1583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1567..1583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1580..1602)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1591..1607
                     /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(1615..1621,1622..1639,1640..1645))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1660..1681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1798..1815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1969..2557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2049..2068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2728..3519,3520..3588))
                     /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     3351..3370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3589..3693)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3768..3787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3959..4338
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4339..4542
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4493..4513
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4587..4606
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4587..4605
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4620..4629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7086..7109
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(7145..7162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7151..7375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7421..7830,1..19)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7508..7527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7718..7739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
       61 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
      121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      181 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
      241 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
      301 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
      361 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
      421 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      481 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      541 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      601 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
      661 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
      721 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
      781 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
      841 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
      901 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
      961 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     1021 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     1081 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     1141 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     1201 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     1261 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     1321 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1381 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1441 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1501 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     1561 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1621 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1681 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1741 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1801 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1861 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1921 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1981 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2041 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2101 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2161 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2221 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2281 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2341 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2401 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2461 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2521 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2581 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2641 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2701 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2761 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2821 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2881 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2941 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3001 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3061 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3121 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3181 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3241 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3301 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3361 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3421 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3481 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3541 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3601 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3661 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3721 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     3781 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     3841 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     3901 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     3961 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4021 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4081 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4141 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4201 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4261 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4321 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     4381 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     4441 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     4501 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
     4561 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggg
     4621 ccaccatggc tctcgtcttc agtgccctgc tgttactggg gctgtgtgga aagatctctt
     4681 cagaaggtca gcctgcattc cataacactc ctggggctat gaattatgaa ttgcctacca
     4741 ccaaatatga gacccaagat accttcaatg ctgggattgt tggccctctc tacaaaatgg
     4801 tgcacatctt cctcaacgtg gtccagccga atgacttccc tctagatttg atcaaaaaac
     4861 tcatacagaa caagaacttt gacatctcag ttgattccaa ggagccagaa atcatagtct
     4921 tggctctgaa gattgccctc tatgagatcg gagtccttat ctgcgccatc ctgggactgc
     4981 tgttcattat cctcatgcct ctggtgggct gcttcttttg tatgtgccgt tgctgcaaca
     5041 aatgcggcgg agagatgcac cagcggcaga agcagaatgc gccatgcagg aggaagtgct
     5101 tgggcctctc cctcctggtg atttgtctgc tcatgagcct tggcattata tatggctttg
     5161 tggctaacca gcagaccagg actcggatca aagggaccca gaaactggca aagagcaatt
     5221 tcagagactt tcaaacactc ctgactgaaa caccaaagca aattgactat gtagtggagc
     5281 agtacaccaa caccaagaac aaggcattct cagacctgga tggcatcggc tccgtgctgg
     5341 gaggcagaat aaaggaccaa ctaaaaccca aagtaactcc tgtcctcgaa gagattaagg
     5401 ccatggcgac agccatcaaa cagaccaagg atgccctgca gaacatgagc agcagcctga
     5461 aaagtctcca agatgcagcc acccagctca ataccaacct gagctctgtg agaaacagca
     5521 tcgagaattc gctcagcagc agtgactgta cctcagatcc agccagcaag atctgcgata
     5581 gcatcagacc aagcctaagc agtctgggga gcagcctcaa ttcaagtcag ctcccatcag
     5641 tggatagaga actcaacact gttactgaag tcgacaaaac tgatctggag agcctcgtca
     5701 aaagggggta tacgacaatt gatgaaatac ccaatacaat acaaaaccaa actgtggatg
     5761 tcatcaaaga cgtcaaaaat accttggact ccattagctc caacattaag gacatgagcc
     5821 aaagtattcc tattgaggat atgctgttac aggtctccca ttaccttaat aacagcaaca
     5881 gatacttaaa ccaggagctg cccaagctgg aagaatatga ctcgtactgg tggctgggtg
     5941 gcttgattgt ctgctttctg ctgactctca ttgtgacctt ctttttcctg ggcttgctgt
     6001 gtggtgtgtt tggctatgac aagcatgcca ccccaactag aagaggctgt gtgtccaaca
     6061 ctggaggcat cttcctcatg gctggggttg gattcggctt ccttttttgc tggatattga
     6121 tgatccttgt ggttcttacg tttgttgttg gtgcaaatgt ggaaaagttg ctctgcgaac
     6181 cttatgaaaa caagaaatta ttacaggttt tggacactcc ctatctgctc aaggaacaat
     6241 ggcaatttta tctttctggc atgctattca ataacccaga cattaacatg acctttgagc
     6301 aagtctacag ggattgcaaa agaggtcgag gtatatatgc tgcttttcag cttgagaatg
     6361 tcgtcaacgt cagtgatcat ttcaacattg accagatttc tgaaaacata aatacggagt
     6421 tggaaaacct gaatgtgaac attgatagca ttgaactgtt ggataacaca ggaaggaaga
     6481 gcctcgagga ctttgcacat tctgggatag atacaatcga ttattccaca tacttgaagg
     6541 agactgagaa atcccctact gaagtgaatc tgctgacatt tgcctctacc ctggaagcaa
     6601 aagcaaacca gttgcctgaa ggaaagctga aacaggcctt cttactggat gtacagaata
     6661 taagagccat ccaccaggtc tggaccctcc agcaaacaag caacaagttg ccggagaaag
     6721 tgaagaagat ccttgcctct ttggactctg ttcagcattt cctcaccaat aacgtttccc
     6781 tcatcgttat cggggaaacg aagaagtttg ggaaaacaat actaggctac tttgaacatt
     6841 atctgcactg ggtcttttat gccatcacag agaagatgac atcctgcaaa cccatggcca
     6901 ccgcgatgga ctctgctgtt aatggcattc tgtgtggcta tgttgcggac cctctgaatt
     6961 tgttctggtt cggcataggg aaagccacgg tgctcttact tccggctgta atcattgcta
     7021 tcaagctggc caagtactat cgcaggatgg attcagagga tgtatacgac gacccgtctc
     7081 gatacgacta caaagacgat gacgacaagt gagtctagag ggcccgttta aacccgctga
     7141 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     7201 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     7261 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     7321 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     7381 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     7441 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     7501 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7561 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7621 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7681 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7741 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     7801 gcctattggt taaaaaatga gctgatttaa
//
