pCMV-mEGFP-iPAK4

LOCUS       pCMV-mEGFP-iPAK4             6667 bp DNA     circular SYN 23-APR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  AUTHORS   Koushik SV, Chen H, Thaler C, Puhl HL, Vogel SS
  TITLE     Cerulean, Venus, and VenusY67C FRET reference standards.
  JOURNAL   Biophys J. 2006 Dec 15. 91(12):L99-L101.
  PUBMED    17040988
REFERENCE   2  (bases 1 to 6667)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 13, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biophys J. 
            2006 Dec 15. 91(12):L99-L101."
FEATURES             Location/Qualifiers
     source          1..6667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1396..2472
                     /mol_type="genomic DNA"
                     /organism="."
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    591..671
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     651..699
                     /label=pUBC-mChilada-f
     primer_bind     652..697
                     /label=pUBC-mCherry-F
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             679..1395
                     /codon_start=1
                     /label=mEGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1281..1315)
                     /label=mEGFP-Qch-R
     primer_bind     1281..1315
                     /label=mEGFP-Qch-F
     misc_feature    1297..1299
                     /label=A to K for mEGFP
     primer_bind     1374..1421
                     /label=GFP-iPAK4-F
     primer_bind     1374..1421
                     /label=Primer 1a
     CDS             1408..1521
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1528..2469
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1621..1639)
                     /label=PAK4-R
     primer_bind     complement(2451..2491)
                     /label=NotI-PAK4-R
     polyA_signal    2594..2715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2631..2650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2685..2704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2722..3177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2859..2880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3071..3090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3204..3308
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3292..3336
                     /label=Primer 1
     CDS             3309..4169
                     /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     complement(4143..4187)
                     /label=Primer 3
     promoter        4171..4528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4194..4214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     4374..4386
                     /label=mCherry-Flag-R
     rep_origin      4379..4514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4441..4460
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4464..4475)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4563..5357
                     /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(4617..4636)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5227..5246
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5545..5564)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5589..5636
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5965..6553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6454..6473
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      841 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1081 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1141 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1201 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1261 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1381 gacgagctgt acaagtcagg tggatccgaa gcagaggact ggacagcagc cctactgaac
     1441 aggggtcgca gtcgccagcc cctggtacta ggggacaatt gctttgctga cttggtgcac
     1501 aactggatgg agctgcctga ggaattcccc gccgcccctg ctgttcctgg gccccctggc
     1561 ccccgctcac cacagcggga gccacagcga gtatcccatg agcagttccg ggctgccctg
     1621 cagctggtgg tggacccagg cgacccccgc tcctacctgg acaacttcat caagattggc
     1681 gagggctcca cgggcatcgt gtgcatcgcc accgtgcgca gctcgggcaa gctggtggcc
     1741 gtcaagaaga tggacctgcg caagcagcag aggcgcgagc tgctcttcaa cgaggtggta
     1801 atcatgaggg actaccagca cgagaatgtg gtggagatgt acaacagcta cctggtgggg
     1861 gacgagctct gggtggtcat ggagttcctg gaaggaggcg ccctcaccga catcgtcacc
     1921 cacaccagga tgaacgagga gcagatcgca gccgtgtgcc ttgcagtgct gcaggccctg
     1981 tcggtgctcc acgcccaggg cgtcatccac cgggacatca agagcgactc gatcctgctg
     2041 acccatgatg gcagggtgaa gctgtcagac tttgggttct gcgcccaggt gagcaaggaa
     2101 gtgccccgaa ggaagtcgct ggtcggcacg ccctactgga tggccccaga gctcatctcc
     2161 cgccttccct acgggccaga ggtagacatc tggtcgctgg ggataatggt gattgagatg
     2221 gtggacggag agccccccta cttcaacgag ccacccctca aagccatgaa gatgattcgg
     2281 gacaacctgc caccccgact gaagaacctg cacaaggtgt cgccatccct gaagggcttc
     2341 ctggaccgcc tgctggtgcg agaccctgcc cagcgggcca cggcagccga gctgctgaag
     2401 cacccattcc tggccaaggc agggccgcct gccagcatcg tgcccctcat gcgccagaac
     2461 cgcaccagat gaagcggccg cgactctaga cataatcagc cataccacat ttgtagaggt
     2521 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2581 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2701 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2761 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2821 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2881 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2941 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3001 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3061 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3121 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3181 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3241 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3301 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3361 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3421 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3481 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3541 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3601 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3661 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3721 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3781 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3841 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3901 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3961 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4021 gtggctctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4081 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4141 taggtgcctc actgattaag cattggtaac ctgaggcgga aagaaccagc tgtggaatgt
     4201 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4321 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4381 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4441 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4501 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4561 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4621 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4681 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4741 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4801 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4861 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4921 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4981 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5041 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5101 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5161 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5221 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5281 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5341 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5401 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5461 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5521 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5581 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5641 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5701 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5761 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5821 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5881 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5941 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6001 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6061 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6121 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6181 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6241 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6301 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6361 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6421 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6481 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6541 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6601 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6661 catgcat
//