LOCUS       sequence-247072-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  AUTHORS   Hoppe AD, Swanson JA
  TITLE     Cdc42, Rac1, and Rac2 display distinct patterns of activation during
            phagocytosis.
  JOURNAL   Mol Biol Cell 2004 Aug;15(8):3509-19. Epub 2004 May 28.
  PUBMED    15169870
REFERENCE   2  (bases 1 to 5304)
  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: "Mol Biol 
            Cell"; date: "2004-08"; volume: "15(8)"; pages: "3509-19. Epub 2004 
            May 28"
COMMENT     Sequence Label: YFP-Rac1
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(155..743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(235..254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1072..1119
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1144..1163
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1351..2145)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1351..2145)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1462..1481)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2072..2091
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2180..2537)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2194..2329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2346..2537
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2494..2514
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2539..2643)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2670..3125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2757..2776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2967..2988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3132..3253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3143..3162)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3197..3216
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3422..4000)
                     /codon_start=1
                     /product="human Rac family small GTPase 1"
                     /label=Rac1
                     /translation="MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH
                     CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR
                     GLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     CDS             complement(join(4016..4726,4727..4729,4730..4732))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4016..4726,4727..4729,4730..4732))
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4058..4079)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4407..4426
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4666..4687
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4729..4738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4777..4980)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4806..4826)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4981..5284
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt
       61 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      121 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      181 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      241 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      301 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      361 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      421 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      481 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      541 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      601 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      661 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      721 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      781 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      841 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac
      901 ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg
      961 aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc
     1021 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
     1081 acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     1141 gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca
     1201 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     1261 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     1321 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     1381 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     1441 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     1501 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     1561 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc
     1621 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     1681 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     1741 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     1801 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     1861 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     1921 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc
     1981 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     2041 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     2101 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     2161 tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     2221 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     2281 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     2341 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     2401 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     2461 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     2521 ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2641 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     2701 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2761 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2821 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2881 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2941 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3001 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3061 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3121 aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3181 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3241 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3301 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3361 gttatctaga tccggtggat cccgggcccg cggtaccgtc gactgcagaa ttcgaagctt
     3421 gtcacaacag caggcatttt ctcttcctct tcttcacggg aggcgggcag aggactgctc
     3481 ggatcgcttc gtcaaacact gtcttgaggc ctcgctgtgt gagcgccgag cactccaggt
     3541 attttacagc accaatctcc ttagccatgg ctagaccctg cggataggtg atgggagtca
     3601 gcttcttctc cttcagtttc tcgatcgtgt ctttatcatc cctaagatca agtttagttc
     3661 ccactaggat gatgggagtg ttgggacagt ggtgccgcac ctcaggatac cactttgcac
     3721 ggacattttc aaatgatgca ggactcacaa gggaaaagca aattaagaac acatctgttt
     3781 gcggatagga tagggggcgt aatctgtcat aatcttcttg tccagctgta tcccataagc
     3841 ccagattcac cggttttcca tctaccataa cattggcaga ataattgtca aagacagtag
     3901 ggatatattc tccaggaaat gcattggttg tgtaactgat cagtaggcaa gttttaccta
     3961 cagctccgtc tcccaccacc acacacttga tggcctgcat agatctgagt ccggacttgt
     4021 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     4081 gatcgcgctt ctcgttgggg tctttgctca gtttggactg gtagctcagg tagtggttgt
     4141 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     4201 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     4261 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     4321 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     4381 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     4441 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     4501 ggtcggggta gcgggcgaag cacatcaggc cgtagccgaa ggtggtcacg agggtgggcc
     4561 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     4621 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     4681 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     4741 ccggtagcgc tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca
     4801 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     4861 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     4921 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     4981 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     5041 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     5101 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     5161 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     5221 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     5281 aacgcggaac tccatatatg ggct
//
