LOCUS       sequence-384596-        5813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for GFP tagged NHERF1. PDZ1 GYGF>GYAA 
            motif mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5813)
  AUTHORS   Garbett D, LaLonde DP, Bretscher A
  TITLE     The scaffolding protein EBP50 regulates microvillar assembly in a 
            phosphorylation-dependent manner.
  JOURNAL   J Cell Biol. 2010 Oct 18;191(2):397-413. doi: 10.1083/jcb.201004115.
            Epub 2010 Oct 11.
  PUBMED    20937695
REFERENCE   2  (bases 1 to 5813)
  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"; doi: 
            "10.1083/jcb.201004115"; journalName: "J Cell Biol"; date: 
            "2010-10-18- 18"; volume: "191"; issue: "2"; pages: "397-413"
COMMENT     Sequence Label: GFP-NHERF1 PDZ1*
FEATURES             Location/Qualifiers
     source          1..5813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        301..504
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     455..475
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(549..551,552..554,555..1265)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(549..551,552..554,555..1265)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(594..615)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(855..874)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1202..1223
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2537..2658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2574..2593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2628..2647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2665..3120)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2802..2823)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3014..3033
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3147..3251
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3253..3610
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3253..3444
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3276..3296)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3461..3596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3645..4439
                     /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             3645..4439
                     /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(3699..3718)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4309..4328
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4627..4646)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4671..4718
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5047..5635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5536..5555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5810..5813,1..300)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
       61 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      121 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      181 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      241 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      301 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      361 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      421 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      481 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      541 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      601 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      661 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      721 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      781 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      841 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      901 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      961 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1021 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1081 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1141 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1201 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1261 acaagtccgg ccggactcag atcctgatga gcgcggacgc agcggccggg gcgcccctgc
     1321 cccggctctg ctgcctggag aagggtccga acggctacgc cgcccacctg cacggggaga
     1381 agggcaagtt gggccagtac atccggctgg tggagcccgg ctcgccggcc gagaaggcgg
     1441 ggctgctggc gggggaccgg ctggtggagg tgaacgggga gaacgtggaa aaggagaccc
     1501 accagcaggt ggtgagccgc atccgcgccg cactcaacgc cgtgcgcctg ctggtggtcg
     1561 accccgagac ggacgagcag ctgcagaagc tcggcgtcca ggtccgagag gagctgctgc
     1621 gcgcccagga agcgccgggg caggccgagc cgccggccgc cgccgaggtg cagggggctg
     1681 gcaacgaaaa tgagcctcgc gaggccgaca agagccaccc ggagcagcgc gagcttcggc
     1741 ctcggctctg taccatgaag aagggcccca gtggctatgg cttcaacctg cacagcgaca
     1801 agtccaagcc aggccagttc atccggtcag tggacccaga ctccccggct gaggcttcag
     1861 ggctccgggc ccaggatcgc attgtggagg tgaacggggt ctgcatggag gggaagcagc
     1921 atggggacgt ggtgtccgcc atcagggctg gcggggacga gaccaagctg ctggtggtgg
     1981 acagggaaac tgacgagttc ttcaagaaat gcagagtgat cccatctcag gagcacctga
     2041 atggtcccct gcctgtgccc ttcaccaatg gggagataca gaaggagaac agtcgtgaag
     2101 ccctggcaga ggcagccttg gagagcccca ggccagccct ggtgagatcc gcctccagtg
     2161 acaccagcga ggagctgaat tcccaagaca gccccccaaa acaggactcc acagcgccct
     2221 cgtctacctc ctcctccgac cccatcctag acttcaacat ctccctggcc atggccaaag
     2281 agagggccca ccagaaacgc agcagcaaac gggccccgca gatggactgg agcaagaaaa
     2341 acgaactctt cagcaacctc tgagccgggt cgaggggaat tctgcagtcg acggtaccgc
     2401 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     2461 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2521 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2581 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2641 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2701 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2761 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2821 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2881 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2941 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3001 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3061 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3241 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3301 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3361 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3421 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3481 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3541 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3601 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3661 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3721 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3781 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3841 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3901 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3961 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4021 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4081 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4141 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4201 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4261 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4321 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4381 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4441 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4501 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4561 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4621 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4681 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4741 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4801 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4861 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4921 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4981 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5041 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5101 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5161 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5221 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5281 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5341 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5401 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5461 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5521 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5581 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5641 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5701 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5761 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gtt
//
