LOCUS       sequence-362470-        7832 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 273-287..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7832)
  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 7832)
  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 Del EC2
FEATURES             Location/Qualifiers
     source          1..7832
                     /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      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3364..3387
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3423..3440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3429..3653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3699..4127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3786..3805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3996..4017
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4136..4156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4141..4470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4321..4456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4383..4402
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4537..5331
                     /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(4591..4610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5201..5220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5505..5626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5542..5561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5596..5615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5675..5691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5675..5691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5688..5710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5699..5715
                     /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(5723..5729,5730..5747,5748..5753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5768..5789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5906..5923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6077..6665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6157..6176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6836..7627,7628..7696))
                     /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     7459..7478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7697..7801)
                     /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 gctggccacc
      901 atggctctcg tcttcagtgc cctgctgtta ctggggctgt gtggaaagat ctcttcagaa
      961 ggtcagcctg cattccataa cactcctggg gctatgaatt atgaattgcc taccaccaaa
     1021 tatgagaccc aagatacctt caatgctggg attgttggcc ctctctacaa aatggtgcac
     1081 atcttcctca acgtggtcca gccgaatgac ttccctctag atttgatcaa aaaactcata
     1141 cagaacaaga actttgacat ctcagttgat tccaaggagc cagaaatcat agtcttggct
     1201 ctgaagattg ccctctatga gatcggagtc cttatctgcg ccatcctggg actgctgttc
     1261 attatcctca tgcctctggt gggctgcttc ttttgtatgt gccgttgctg caacaaatgc
     1321 ggcggagaga tgcaccagcg gcagaagcag aatgcgccat gcaggaggaa gtgcttgggc
     1381 ctctccctcc tggtgatttg tctgctcatg agccttggca ttatatatgg ctttgtggct
     1441 aaccagcaga ccaggactcg gatcaaaggg acccagaaac tggcaaagag caatttcaga
     1501 gactttcaaa cactcctgac tgaaacacca aagcaaattg actatgtagt ggagcagtac
     1561 accaacacca agaacaaggc attctcagac ctggatggca tcggctccgt gctgggaggc
     1621 agaataaagg accaactaaa acccaaagta actcctgtcc tcgaagagat taaggccatg
     1681 gcgacagcca tcaaacagac caaggatgcc acccagctca ataccaacct gagctctgtg
     1741 agaaacagca tcgagaattc gctcagcagc agtgactgta cctcagatcc agccagcaag
     1801 atctgcgata gcatcagacc aagcctaagc agtctgggga gcagcctcaa ttcaagtcag
     1861 ctcccatcag tggatagaga actcaacact gttactgaag tcgacaaaac tgatctggag
     1921 agcctcgtca aaagggggta tacgacaatt gatgaaatac ccaatacaat acaaaaccaa
     1981 actgtggatg tcatcaaaga cgtcaaaaat accttggact ccattagctc caacattaag
     2041 gacatgagcc aaagtattcc tattgaggat atgctgttac aggtctccca ttaccttaat
     2101 aacagcaaca gatacttaaa ccaggagctg cccaagctgg aagaatatga ctcgtactgg
     2161 tggctgggtg gcttgattgt ctgctttctg ctgactctca ttgtgacctt ctttttcctg
     2221 ggcttgctgt gtggtgtgtt tggctatgac aagcatgcca ccccaactag aagaggctgt
     2281 gtgtccaaca ctggaggcat cttcctcatg gctggggttg gattcggctt ccttttttgc
     2341 tggatattga tgatccttgt ggttcttacg tttgttgttg gtgcaaatgt ggaaaagttg
     2401 ctctgcgaac cttatgaaaa caagaaatta ttacaggttt tggacactcc ctatctgctc
     2461 aaggaacaat ggcaatttta tctttctggc atgctattca ataacccaga cattaacatg
     2521 acctttgagc aagtctacag ggattgcaaa agaggtcgag gtatatatgc tgcttttcag
     2581 cttgagaatg tcgtcaacgt cagtgatcat ttcaacattg accagatttc tgaaaacata
     2641 aatacggagt tggaaaacct gaatgtgaac attgatagca ttgaactgtt ggataacaca
     2701 ggaaggaaga gcctcgagga ctttgcacat tctgggatag atacaatcga ttattccaca
     2761 tacttgaagg agactgagaa atcccctact gaagtgaatc tgctgacatt tgcctctacc
     2821 ctggaagcaa aagcaaacca gttgcctgaa ggaaagctga aacaggcctt cttactggat
     2881 gtacagaata taagagccat ccaccagcat ctcctccctc ctgtgcagca atcactgaat
     2941 acgttaagac aaagtgtctg gaccctccag caaacaagca acaagttgcc ggagaaagtg
     3001 aagaagatcc ttgcctcttt ggactctgtt cagcatttcc tcaccaataa cgtttccctc
     3061 atcgttatcg gggaaacgaa gaagtttggg aaaacaatac taggctactt tgaacattat
     3121 ctgcactggg tcttttatgc catcacagag aagatgacat cctgcaaacc catggccacc
     3181 gcgatggact ctgctgttaa tggcattctg tgtggctatg ttgcggaccc tctgaatttg
     3241 ttctggttcg gcatagggaa agccacggtg ctcttacttc cggctgtaat cattgctatc
     3301 aagctggcca agtactatcg caggatggat tcagaggatg tatacgacga cccgtctcga
     3361 tacgactaca aagacgatga cgacaagtga gtctagaggg cccgtttaaa cccgctgatc
     3421 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3601 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3661 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3721 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3781 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3841 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3901 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3961 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4021 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4081 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4141 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4261 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4381 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4441 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4501 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4561 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4621 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4681 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4741 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4801 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4861 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4921 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4981 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5041 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5101 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5161 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5221 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5281 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5341 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5401 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5461 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5521 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5581 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5641 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7801 gcacatttcc ccgaaaagtg ccacctgacg tc
//
