pCMV-mNeonGreen-iPAK4

LOCUS       pCMV-mNeonGreen-iPAK4             6658 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 6658)
  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 6658)
  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..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1387..2463
                     /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
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             679..1386
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     primer_bind     complement(1205..1245)
                     /label=mNeonGreen_G183T-R
     primer_bind     1228..1252
                     /label=Vector_mNeonGreen-F
     primer_bind     1366..1412
                     /label=GFP-iPAK4-F
     primer_bind     complement(1366..1412)
                     /label=XFP-iPAK4-R
     CDS             1399..1512
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1519..2460
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1612..1630)
                     /label=PAK4-R
     primer_bind     complement(2442..2482)
                     /label=NotI-PAK4-R
     polyA_signal    2585..2706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2622..2641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2676..2695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2713..3168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2850..2871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3062..3081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3195..3299
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3283..3327
                     /label=Primer 1
     CDS             3300..4160
                     /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(4134..4178)
                     /label=Primer 3
     promoter        4162..4519
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4185..4205)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4370..4505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4432..4451
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4455..4466)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4554..5348
                     /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(4608..4627)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5218..5237
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5536..5555)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5580..5627
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5956..6544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6445..6464
                     /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 ggcgaggagg ataacatggc ctctctccca
      721 gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt
      781 cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt
      841 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
      901 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggatac
      961 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     1021 acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct
     1081 gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact
     1141 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac gggaaatggc
     1201 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     1261 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc
     1321 gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg
     1381 tacaagtcag gtggatccga agcagaggac tggacagcag ccctactgaa caggggtcgc
     1441 agtcgccagc ccctggtact aggggacaat tgctttgctg acttggtgca caactggatg
     1501 gagctgcctg aggaattccc cgccgcccct gctgttcctg ggccccctgg cccccgctca
     1561 ccacagcggg agccacagcg agtatcccat gagcagttcc gggctgccct gcagctggtg
     1621 gtggacccag gcgacccccg ctcctacctg gacaacttca tcaagattgg cgagggctcc
     1681 acgggcatcg tgtgcatcgc caccgtgcgc agctcgggca agctggtggc cgtcaagaag
     1741 atggacctgc gcaagcagca gaggcgcgag ctgctcttca acgaggtggt aatcatgagg
     1801 gactaccagc acgagaatgt ggtggagatg tacaacagct acctggtggg ggacgagctc
     1861 tgggtggtca tggagttcct ggaaggaggc gccctcaccg acatcgtcac ccacaccagg
     1921 atgaacgagg agcagatcgc agccgtgtgc cttgcagtgc tgcaggccct gtcggtgctc
     1981 cacgcccagg gcgtcatcca ccgggacatc aagagcgact cgatcctgct gacccatgat
     2041 ggcagggtga agctgtcaga ctttgggttc tgcgcccagg tgagcaagga agtgccccga
     2101 aggaagtcgc tggtcggcac gccctactgg atggccccag agctcatctc ccgccttccc
     2161 tacgggccag aggtagacat ctggtcgctg gggataatgg tgattgagat ggtggacgga
     2221 gagcccccct acttcaacga gccacccctc aaagccatga agatgattcg ggacaacctg
     2281 ccaccccgac tgaagaacct gcacaaggtg tcgccatccc tgaagggctt cctggaccgc
     2341 ctgctggtgc gagaccctgc ccagcgggcc acggcagccg agctgctgaa gcacccattc
     2401 ctggccaagg cagggccgcc tgccagcatc gtgcccctca tgcgccagaa ccgcaccaga
     2461 tgaagcggcc gcgactctag acataatcag ccataccaca tttgtagagg ttttacttgc
     2521 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2581 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2641 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2701 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3121 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3181 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3241 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3301 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3361 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3421 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3481 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3541 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3601 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3661 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3721 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3781 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3841 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3901 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3961 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggctctc
     4021 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4081 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4141 cactgattaa gcattggtaa cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4201 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4261 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4321 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4381 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4441 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4501 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4561 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4621 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4681 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4741 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4801 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4861 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4921 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4981 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5041 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5101 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5161 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5221 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5281 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5341 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5401 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5461 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5521 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5581 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5641 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5701 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5761 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5821 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5881 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6061 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6601 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//