LOCUS       sequence-456539-        6079 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 6079)
  AUTHORS   Abell AN, DeCathelineau AM, Weed SA, Ambruso DR, Riches DW, Johnson 
            GL
  TITLE     Rac2D57N, a dominant inhibitory Rac2 mutant that inhibits p38 kinase
            signaling and prevents surface ruffling in bone-marrow-derived 
            macrophages.
  JOURNAL   J Cell Sci. 2004 Jan 15. 117(Pt 2):243-55.
  PUBMED    14676277
REFERENCE   2  (bases 1 to 6079)
  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: "J Cell Sci.
            2004 Jan 15. 117(Pt 2):243-55."
COMMENT     Sequence Label: pcDNA3-Flag Rac2
FEATURES             Location/Qualifiers
     source          1..6079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     595..614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(833..937)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1012..1031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1203..1582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1583..1786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1737..1757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1831..1850
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1831..1849
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1862..1871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1871..1894
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2600..2618)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2601..2618)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2638..2655)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2644..2868
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2914..3342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3001..3020)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3211..3232
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3351..3371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3356..3685
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3536..3671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3598..3617
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3752..4546
                     /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(3806..3825)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4416..4435
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4720..4841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4757..4776)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4811..4830
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4890..4906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4890..4906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4903..4925)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4914..4930
                     /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(4938..4944,4945..4962,4963..4968))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4983..5004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5121..5138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5292..5880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5372..5391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6051..6079,1..763,764..832))
                     /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"
ORIGIN
        1 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
       61 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      961 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     1021 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     1081 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     1141 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     1201 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1261 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1561 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     1621 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1681 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1741 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
     1801 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     1861 tgccaccatg gactacaagg acgacgatga caagggacct aagcttggta ccgagctcgg
     1921 atccggtacc atgcaggcca tcaagtgtgt ggtggtggga gatggggccg tgggcaagac
     1981 ctgccttctc atcagctaca ccaccaacgc ctttcccgga gagtacatcc ccaccgtgtt
     2041 tgacaactat tcagccaatg tgatggtgga cagcaagcca gtgaacctgg ggctgtggga
     2101 cactgctggg caggaggact acgaccgtct ccggccgctc tcctatccac agacggacgt
     2161 cttcctcatc tgcttctccc tcgtcagccc agcctcttat gagaacgtcc gcgccaagtg
     2221 gttcccagaa gtgcggcacc actgccccag cacacccatc atcctggtgg gcaccaagct
     2281 ggacctgcgg gacgacaagg acaccatcga gaaactgaag gagaagaagc tggctcccat
     2341 cacctacccg cagggcctgg cactggccaa ggagattgac tcggtgaaat acctggagtg
     2401 ctcagctctc acccagagag gcctgaaaac cgtgttcgac gaggccatcc gggccgtgct
     2461 gtgccctcag cccacgcggc agcagaagcg cgcctgcagc ctcctctagg atccactagt
     2521 aacggccgcc agtgtgctgg aattctgcag atatccatca cactggcggc cgctcgagca
     2581 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     2641 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2701 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2761 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2821 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2881 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2941 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3001 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3061 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3121 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3181 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3241 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3301 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3361 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3421 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3481 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3541 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3601 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3661 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3721 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3781 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3841 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3901 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3961 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4021 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4081 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4141 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4201 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4261 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4321 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4381 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4441 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4501 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4561 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4621 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4681 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4741 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4801 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4861 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4921 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4981 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5041 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5101 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5161 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5221 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5281 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5341 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5401 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5461 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5521 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5581 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5641 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5701 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5761 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5821 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5881 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5941 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6001 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6061 ttaatcagtg aggcaccta
//
