LOCUS       sequence-142743-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CDC42EP1-V5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  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 6648)
  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 CDC42EP1 S192A
FEATURES             Location/Qualifiers
     source          1..6648
                     /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(2088..2112)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2138..2179
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2219..2238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2263..2311
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2513..2941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2600..2619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2810..2831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2950..2970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2955..3284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3135..3270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3197..3216
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3351..4145
                     /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(3405..3424)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4015..4034
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4321..4442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4358..4377)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4412..4431
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4491..4507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4491..4507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4504..4526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4515..4531
                     /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(4539..4545,4546..4563,4564..4569))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4584..4605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4722..4739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4893..5481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4973..4992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5652..6443,6444..6512))
                     /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     6275..6294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6513..6617)
                     /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 tggcatgccc ggcccccagg ggggcagagg
      961 cgccgccacc atgagcctgg gcaagctctc gcctgtgggc tgggtgtcca gttcacaggg
     1021 aaagaggcgg ctgactgcag acatgatcag ccacccactc ggggacttcc gccacaccat
     1081 gcatgtgggc cgtggcgggg atgtcttcgg ggacacgtcc ttcctcagca accacggtgg
     1141 cagctccggg agcacccatc gctcaccccg cagcttcctg gccaagaagc tgcagctggt
     1201 gcggagggtg ggggcgcccc cccggaggat ggcatctccc cctgcaccct ccccggctcc
     1261 accggccatc tcccccatca tcaagaacgc catctccctg ccccagctca accaggccgc
     1321 ctacgacagc ctcgtggttg gcaagctcag cttcgacagc agccccacca gctccacgga
     1381 cggccactcc agctacggcc tggactctgg gttctgcacc atctcccgcc tgccccgctc
     1441 ggaaaagccg catgaccgag accgggatgg ttccttcccc tctgagcccg ggcttcgccg
     1501 ctctgacgct ctcttgtcct tccgcctgga cctcgacctt gggccctcac tcctcagcga
     1561 gctgctaggg gtcatgagcc tgccagaagc ccctgcagct gagactccag cccccgctgc
     1621 aaacccccca gcccctactg caaaccccac gggtcctgct gcaaaccccc cagccactac
     1681 tgcaaacccc ccagcacctg ccgcaacccc cacgggtcct gctgcaaatc ccccagcccc
     1741 tgccgcaagc tccacacccc atggacactg tcccaatggg gtaacagctg ggttgggccc
     1801 agtggctgag gtgaagtcca gcccagtggg agggggtccc cgaggacctg ctggccctgc
     1861 cctcggcagg cactggggag caggctggga tggcggccac cactacccag agatggatgc
     1921 gcggcaggag cgggtggagg tgctgcccca agcccgggcc tcctgggaga gcctggacga
     1981 agagtggagg gcgccccagg caggcagcag gaccccagtg cccagcacag tgcaagcaaa
     2041 cacctttgaa tttgcggatg ctgaggagga tgatgaggtc aaggtgtgcc caactttctt
     2101 gtacaaagtg gttgatctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     2161 cctcggtctc gattctacgc gtaccggtta gtaatgagtt taaacggggg aggctaactg
     2221 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     2281 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     2341 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     2401 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     2461 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     2521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2641 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     2701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2881 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2941 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3061 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3241 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3301 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3361 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3421 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3481 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3541 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3601 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3661 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3721 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3781 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3841 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3901 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3961 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4021 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4081 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4141 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     4201 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4261 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4321 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4381 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4441 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4501 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4561 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4621 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4681 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5641 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5701 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5761 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5821 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5881 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5941 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6001 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6061 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6121 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6181 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6241 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6301 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6361 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6421 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6481 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6541 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6601 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
