LOCUS       sequence-142810-        7503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RBM14-V5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7503)
  AUTHORS   Schaffer BE, Levin RS, Hertz NT, Maures TJ, Schoof ML, Hollstein PE,
            Benayoun BA, Banko MR, Shaw RJ, Shokat KM, Brunet A
  TITLE     Identification of AMPK Phosphorylation Sites Reveals a Network of 
            Proteins Involved in Cell Invasion and Facilitates Large-Scale 
            Substrate Prediction.
  JOURNAL   Cell Metab. 2015 Nov 3;22(5):907-21. doi: 
            10.1016/j.cmet.2015.09.009. Epub 2015 Oct 8.
  PUBMED    26456332
REFERENCE   2  (bases 1 to 7503)
  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"; doi: 
            "10.1016/j.cmet.2015.09.009"; journalName: "Cell Metab"; date: 
            "2015-11-3- 3"; volume: "22"; issue: "5"; pages: "907-21"
COMMENT     Sequence Label: pcDNA 3.2/V5-DEST RBM14-V5
FEATURES             Location/Qualifiers
     source          1..7503
                     /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"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2943..2967)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2993..3034
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3074..3093)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3118..3166
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3368..3796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3455..3474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3665..3686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3805..3825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3810..4139
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3990..4125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4206..5000
                     /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(4260..4279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4870..4889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5176..5297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5213..5232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5267..5286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5346..5362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5346..5362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5359..5381)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5370..5386
                     /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(5394..5400,5401..5418,5419..5424))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5439..5460
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5577..5594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5748..6336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5828..5847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6507..7298,7299..7367))
                     /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     7130..7149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7368..7472)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gctggctagt
      901 taagctatca acaagtttgt acaaaaaagt tggcatgaag atattcgtgg gcaacgtcga
      961 cggggcggat acgactccgg aggagctggc agccctcttt gcgccctacg gcacggtcat
     1021 gagctgcgcc gtcatgaaac agttcgcctt cgtgcacatg cgcgagaacg cgggcgcgct
     1081 gcgcgccatc gaagccctgc acggccacga gctgcggccg gggcgcgcgc tcgtggtgga
     1141 gatgtcgcgc ccaaggcctc ttaatacttg gaagattttc gtgggcaatg tgtcggctgc
     1201 atgcacgagc caggaactgc gcagcctctt cgagcgccgc ggacgcgtca tcgagtgtga
     1261 cgtggtgaaa gactacgcgt ttgttcacat ggagaaggaa gcagatgcca aagccgcaat
     1321 cgcgcagctc aacggcaaag aagtgaaggg caagcgcatc aacgtggaac tctccaccaa
     1381 gggtcagaag aaggggcctg gcctggctgt ccagtctggg gacaagacca agaaaccagg
     1441 ggctggggat acggccttcc ctggaactgg tggcttctct gccaccttcg actaccagca
     1501 ggcttttggc aacagcactg gtggctttga tgggcaagcc cgtcagccca caccaccctt
     1561 ctttggtcgc gaccgcagcc ctctgcgccg ttcacctccc cgagcctctt atgtggctcc
     1621 tctgacggcc cagccagcta cctaccgggc ccagccgtcc gtgtcactgg gagctgccta
     1681 cagggcccag ccttctgcct ctttgggtgt tggctatcgg actcagccca tgacagccca
     1741 ggcagcctct taccgcgctc agccctctgt ctcccttggg gcaccataca ggggccagct
     1801 ggctagtcct agctcccagt ctgctgcagc ttcttcactc ggcccatatg gtggagccca
     1861 gccctcagcc tcggcccttt cctcctatgg gggtcaggca gctgcagctt cttcgctcaa
     1921 ctcctatggg gctcagggtt cctcccttgc ctcctatggt aaccagccat cctcttacgg
     1981 cgcccaggct gcctcttcct atggggttcg tgcagctgct tcttcctaca acacccaggg
     2041 agcagcttcc tccttaggct cctacggggc tcaggcagcc tcctatgggg cccagtctgc
     2101 agcctcctca ctagcttatg gagcccaggc agcttcatat aatgcccagc cctcggcctc
     2161 ttacaatgcc cagtctgccc catatgctgc acagcaggct gcttcctact cttcccaacc
     2221 tgctgcctat gtggcacagc cagccacagc tgctgcctat gccagccagc cagcagccta
     2281 cgccgcacaa gccactaccc caatggctgg ctcctatggg gcccagccgg ttgtgcagac
     2341 ccagctgaat agttacgggg cccaagcatc aatgggcctt tcaggctcct atggggctca
     2401 gtcggctgct gcggccactg gctcctatgg tgccgcagca gcctacgggg cccaaccttc
     2461 tgccaccctg gcagctcctt accgcactca gtcatcagcc tcattggctg cttcctatgc
     2521 tgcccagcag catccccagg ctgctgcctc ctaccgcggc cagccaggca atgcctacga
     2581 tggggcaggt cagccgtctg cagcctacct gtccatgtcc cagggggccg ttgccaacgc
     2641 caacagcacc ccgccgccct atgagcgtac ccgcctctcc ccaccccggg ccagctacga
     2701 cgatccctac aaaaaggctg tcgccatgtc gaaaaggtat ggttccgacc ggcgtttagc
     2761 cgagctctct gattaccgcc gtttatcaga gtcgcagctt tcgttccgcc gctcgccgac
     2821 aaagtcctcg ctggattacc gtcgcctgcc cgatgcccat tccgattacg cacgctattc
     2881 gggctcctat aatgattacc tgcgggcggc tcagatgcac tctggctacc agcgccgcat
     2941 gtacccaact ttcttgtaca aagtggttga tctagagggc ccgcggttcg aaggtaagcc
     3001 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggttagtaat gagtttaaac
     3061 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3121 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3181 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3241 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3301 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3361 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3421 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3481 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     3541 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3601 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3661 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3721 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3781 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3901 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3961 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4021 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4081 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4141 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4201 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4261 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4321 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4381 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4441 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4501 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4561 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4621 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4681 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4741 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4801 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4861 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4921 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4981 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5041 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5101 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5161 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5221 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5281 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5341 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5401 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5461 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5521 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5941 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6181 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7501 gtc
//
