pCMV-mScarletI-iPAK4

LOCUS       pCMV-mScarletI-iPAK4                6646 bp DNA     circular SYN 09-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    mVenus N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6646)
  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 6646)
  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..6646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1375..2451
                     /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     646..684
                     /label=BamHI-iPAK4-F
     primer_bind     652..698
                     /label=mScarletI-1P0K-F
     primer_bind     complement(669..695)
                     /label=mScarletI-FLAG-Qch-R
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             679..1374
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(812..830)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1080..1099
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1305..1341)
                     /label=BamHI-iPAK4-F
     primer_bind     1353..1400
                     /label=GFP-iPAK4-F
     primer_bind     complement(1354..1400)
                     /label=XFP-iPAK4-R
     CDS             1387..1500
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1507..2448
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1600..1618)
                     /label=PAK4-R
     primer_bind     complement(2430..2470)
                     /label=NotI-PAK4-R
     polyA_signal    2573..2694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2610..2629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2664..2683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2701..3156)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2838..2859)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3050..3069
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3183..3287
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3271..3315
                     /label=Primer 1
     CDS             3288..4148
                     /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(4122..4166)
                     /label=Primer 3
     promoter        4150..4507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4173..4193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4358..4493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4420..4439
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4443..4454)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4542..5336
                     /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(4596..4615)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5206..5225
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5524..5543)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5568..5615
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5944..6532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6433..6452
                     /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 ggcgaggcag tgatcaagga gttcatgcgg
      721 ttcaaggtgc acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
      781 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      841 ctgcccttct cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc
      901 aagcaccccg ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg
      961 gagcgcgtga tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg
     1021 gaggacggca ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc
     1081 cccgtaatgc agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag
     1141 gacggcgtgc tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac
     1201 ctggcggact tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac
     1261 aacgtcgacc gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag
     1321 tacgaacgct ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtcaggt
     1381 ggatccgaag cagaggactg gacagcagcc ctactgaaca ggggtcgcag tcgccagccc
     1441 ctggtactag gggacaattg ctttgctgac ttggtgcaca actggatgga gctgcctgag
     1501 gaattccccg ccgcccctgc tgttcctggg ccccctggcc cccgctcacc acagcgggag
     1561 ccacagcgag tatcccatga gcagttccgg gctgccctgc agctggtggt ggacccaggc
     1621 gacccccgct cctacctgga caacttcatc aagattggcg agggctccac gggcatcgtg
     1681 tgcatcgcca ccgtgcgcag ctcgggcaag ctggtggccg tcaagaagat ggacctgcgc
     1741 aagcagcaga ggcgcgagct gctcttcaac gaggtggtaa tcatgaggga ctaccagcac
     1801 gagaatgtgg tggagatgta caacagctac ctggtggggg acgagctctg ggtggtcatg
     1861 gagttcctgg aaggaggcgc cctcaccgac atcgtcaccc acaccaggat gaacgaggag
     1921 cagatcgcag ccgtgtgcct tgcagtgctg caggccctgt cggtgctcca cgcccagggc
     1981 gtcatccacc gggacatcaa gagcgactcg atcctgctga cccatgatgg cagggtgaag
     2041 ctgtcagact ttgggttctg cgcccaggtg agcaaggaag tgccccgaag gaagtcgctg
     2101 gtcggcacgc cctactggat ggccccagag ctcatctccc gccttcccta cgggccagag
     2161 gtagacatct ggtcgctggg gataatggtg attgagatgg tggacggaga gcccccctac
     2221 ttcaacgagc cacccctcaa agccatgaag atgattcggg acaacctgcc accccgactg
     2281 aagaacctgc acaaggtgtc gccatccctg aagggcttcc tggaccgcct gctggtgcga
     2341 gaccctgccc agcgggccac ggcagccgag ctgctgaagc acccattcct ggccaaggca
     2401 gggccgcctg ccagcatcgt gcccctcatg cgccagaacc gcaccagatg aagcggccgc
     2461 gactctagac ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2521 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2701 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2761 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2821 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2881 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2941 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3001 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3061 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3121 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3181 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3241 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3301 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3361 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3421 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3481 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3541 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3601 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3661 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3721 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3781 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3841 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3901 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3961 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg
     4021 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4081 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4141 attggtaacc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4201 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4261 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4321 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4381 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4441 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4501 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4561 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4621 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4681 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4741 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4801 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4861 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4921 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4981 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5041 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5101 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5161 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5221 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5281 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5341 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5401 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5461 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5521 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5581 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5641 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5701 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5761 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5821 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5881 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6601 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//