LOCUS       sequence-208851-        5898 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 5898)
  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 5898)
  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: pIRES2-EGFP Rac2
FEATURES             Location/Qualifiers
     source          1..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    602..658
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    join(1250..1824,1825..1827,1828..1836)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1417..1434)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1644..1663
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1837..1839,1840..1842,1843..2556)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1837..1839,1840..1842,1843..2553)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1882..1903)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2143..2162)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2490..2511
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2679..2800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2716..2735)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2770..2789
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2807..3262)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2944..2965)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3156..3175
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3289..3393
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3395..3752
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3395..3586
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3418..3438)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3603..3738
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3787..4581
                     /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             3787..4581
                     /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(3841..3860)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4451..4470
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4769..4788)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4813..4860
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5189..5777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5678..5697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccgg
      661 taccatgcag gccatcaagt gtgtggtggt gggagatggg gccgtgggca agacctgcct
      721 tctcatcagc tacaccacca acgcctttcc cggagagtac atccccaccg tgtttgacaa
      781 ctattcagcc aatgtgatgg tggacagcaa gccagtgaac ctggggctgt gggacactgc
      841 tgggcaggag gactacgacc gtctccggcc gctctcctat ccacagacgg acgtcttcct
      901 catctgcttc tccctcgtca gcccagcctc ttatgagaac gtccgcgcca agtggttccc
      961 agaagtgcgg caccactgcc ccagcacacc catcatcctg gtgggcacca agctggacct
     1021 gcgggacgac aaggacacca tcgagaaact gaaggagaag aagctggctc ccatcaccta
     1081 cccgcagggc ctggcactgg ccaaggagat tgactcggtg aaatacctgg agtgctcagc
     1141 tctcacccag agaggcctga aaaccgtgtt cgacgaggcc atccgggccg tgctgtgccc
     1201 tcagcccacg cggcagcaga agcgcgcctg cagcctcctc taggatccgc ccctctccct
     1261 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1321 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     1381 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     1441 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     1501 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     1561 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     1621 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     1681 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta
     1741 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     1801 tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg
     1861 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1921 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1981 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2041 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2101 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2161 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2221 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2281 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2341 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2401 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2461 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2521 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     2581 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2641 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2701 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2761 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2821 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2881 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2941 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3001 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3061 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3121 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3181 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3241 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3301 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3361 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3421 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3481 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3601 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3661 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3721 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3781 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3841 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3901 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3961 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4021 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4081 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4141 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4201 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4261 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4321 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4381 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4441 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4501 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4561 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4621 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4681 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4741 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4801 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4861 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4921 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4981 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5041 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5101 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5161 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5221 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5281 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5341 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5401 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5461 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5521 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5581 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5641 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5701 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5761 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5821 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5881 ccgccatgca ttagttat
//
