LOCUS       sequence-362466-        7833 bp    DNA     circular SYN 15-APR-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 702-716..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7833)
  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 7833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..7833
                     /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             3365..3388
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3424..3441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3430..3654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3700..4128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3787..3806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3997..4018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4137..4157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4142..4471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4322..4457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4384..4403
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4538..5332
                     /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(4592..4611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5506..5627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5543..5562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5597..5616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5676..5692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5676..5692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5689..5711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5700..5716
                     /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(5724..5730,5731..5748,5749..5754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5769..5790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5907..5924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6078..6666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6158..6177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6837..7628,7629..7697))
                     /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     7460..7479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7698..7802)
                     /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 aagcaaaagc
     2881 aaaccagttg cctgaaggaa agctgaaaca ggccttctta ctggatgtac agaatataag
     2941 agccatccac cagcatctcc tccctcctgt gcagcaatca ctgaatttgc cggagaaagt
     3001 gaagaagatc cttgcctctt tggactctgt tcagcatttc ctcaccaata acgtttccct
     3061 catcgttatc ggggaaacga agaagtttgg gaaaacaata ctaggctact ttgaacatta
     3121 tctgcactgg gtcttttatg ccatcacaga gaagatgaca tcctgcaaac ccatggccac
     3181 cgcgatggac tctgctgtta atggcattct gtgtggctat gttgcggacc ctctgaattt
     3241 gttctggttc ggcataggga aagccacggt gctcttactt ccggctgtaa tcattgctat
     3301 caagctggcc aagtactatc gcaggatgga ttcagaggat gtatacgacg acccgtctcg
     3361 atacgactac aaagacgatg acgacaagtg agtctagagg gcccgtttaa acccgctgat
     3421 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3481 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3541 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3601 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3661 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3901 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4021 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4081 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4141 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4201 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4381 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4441 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4501 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5401 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5461 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5521 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5581 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5641 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5701 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5761 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5821 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5881 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5941 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6001 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6241 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6481 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6721 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6781 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7801 cgcacatttc cccgaaaagt gccacctgac gtc
//
