pCMV-FLAG-iPAK4

LOCUS       pCMV-FLAG-iPAK4             5966 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 5966)
  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 5966)
  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..5966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          679..1779
                     /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     652..708
                     /label=FLAG-iPAK4-F
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      673..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             715..828
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             835..1776
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(928..946)
                     /label=PAK4-R
     primer_bind     1766..1790
                     /label=NotI-IRES-F
     polyA_signal    1893..2014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1930..1949)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1984..2003
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2021..2476)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2158..2179)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2370..2389
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2503..2607
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2591..2635
                     /label=Primer 1
     CDS             2608..3468
                     /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(3442..3486)
                     /label=Primer 3
     promoter        3470..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3493..3513)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3678..3813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3740..3759
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3763..3774)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3862..4656
                     /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(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4844..4863)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4888..4935
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5264..5852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5753..5772
                     /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 ggactacaag gacgacgatg ataagtccgg atccgaagca
      721 gaggactgga cagcagccct actgaacagg ggtcgcagtc gccagcccct ggtactaggg
      781 gacaattgct ttgctgactt ggtgcacaac tggatggagc tgcctgagga attccccgcc
      841 gcccctgctg ttcctgggcc ccctggcccc cgctcaccac agcgggagcc acagcgagta
      901 tcccatgagc agttccgggc tgccctgcag ctggtggtgg acccaggcga cccccgctcc
      961 tacctggaca acttcatcaa gattggcgag ggctccacgg gcatcgtgtg catcgccacc
     1021 gtgcgcagct cgggcaagct ggtggccgtc aagaagatgg acctgcgcaa gcagcagagg
     1081 cgcgagctgc tcttcaacga ggtggtaatc atgagggact accagcacga gaatgtggtg
     1141 gagatgtaca acagctacct ggtgggggac gagctctggg tggtcatgga gttcctggaa
     1201 ggaggcgccc tcaccgacat cgtcacccac accaggatga acgaggagca gatcgcagcc
     1261 gtgtgccttg cagtgctgca ggccctgtcg gtgctccacg cccagggcgt catccaccgg
     1321 gacatcaaga gcgactcgat cctgctgacc catgatggca gggtgaagct gtcagacttt
     1381 gggttctgcg cccaggtgag caaggaagtg ccccgaagga agtcgctggt cggcacgccc
     1441 tactggatgg ccccagagct catctcccgc cttccctacg ggccagaggt agacatctgg
     1501 tcgctgggga taatggtgat tgagatggtg gacggagagc ccccctactt caacgagcca
     1561 cccctcaaag ccatgaagat gattcgggac aacctgccac cccgactgaa gaacctgcac
     1621 aaggtgtcgc catccctgaa gggcttcctg gaccgcctgc tggtgcgaga ccctgcccag
     1681 cgggccacgg cagccgagct gctgaagcac ccattcctgg ccaaggcagg gccgcctgcc
     1741 agcatcgtgc ccctcatgcg ccagaaccgc accagatgaa gcggccgcgc ataatcagcc
     1801 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1861 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1921 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1981 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2041 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2101 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2161 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2221 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2281 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2341 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2401 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2461 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2821 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3121 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3301 aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg gccagatggt
     3361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc tgaggcggaa
     3481 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4801 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4861 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4921 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4981 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5041 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5101 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5161 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5221 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5281 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5341 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5401 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5461 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5521 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5581 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5641 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5701 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5761 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5821 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5881 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5941 tggataaccg tattaccgcc atgcat
//