LOCUS       sequence-362430-        7878 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 
            689 to 694..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7878)
  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 7878)
  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 AA 6
FEATURES             Location/Qualifiers
     source          1..7878
                     /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             3410..3433
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3469..3486)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3475..3699
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3745..4173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3832..3851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4042..4063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4182..4202)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4187..4516
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4367..4502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4429..4448
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4583..5377
                     /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(4637..4656)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5247..5266
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5551..5672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5588..5607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5642..5661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5721..5737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5721..5737)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5734..5756)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5745..5761
                     /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(5769..5775,5776..5793,5794..5799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5814..5835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5952..5969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6123..6711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6203..6222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6882..7673,7674..7742))
                     /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     7505..7524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7743..7847)
                     /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 tccctaaatt tgtgagggtg aggaatacgt taagacaaag
     3001 tgtctggacc ctccagcaaa caagcaacaa gttgccggag aaagtgaaga agatccttgc
     3061 ctctttggac tctgttcagc atttcctcac caataacgtt tccctcatcg ttatcgggga
     3121 aacgaagaag tttgggaaaa caatactagg ctactttgaa cattatctgc actgggtctt
     3181 ttatgccatc acagagaaga tgacatcctg caaacccatg gccaccgcga tggactctgc
     3241 tgttaatggc attctgtgtg gctatgttgc ggaccctctg aatttgttct ggttcggcat
     3301 agggaaagcc acggtgctct tacttccggc tgtaatcatt gctatcaagc tggccaagta
     3361 ctatcgcagg atggattcag aggatgtata cgacgacccg tctcgatacg actacaaaga
     3421 cgatgacgac aagtgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     3481 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3541 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3601 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3661 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3721 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3781 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3841 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3901 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3961 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4021 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4081 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4141 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4201 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4261 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4321 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4381 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4441 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4501 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4561 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4621 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4681 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4741 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4801 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4861 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4921 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4981 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5041 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5101 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5161 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5221 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5281 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5341 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5401 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5461 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5521 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5581 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5641 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5701 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5761 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5821 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5881 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5941 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6001 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6061 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6121 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6181 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6241 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6301 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6361 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6421 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6481 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6541 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6601 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6661 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6721 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6781 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6841 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6901 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6961 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7021 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7081 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7141 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7201 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7261 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7321 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7381 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7441 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7501 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7561 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7621 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7681 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7741 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7801 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7861 aaagtgccac ctgacgtc
//
