LOCUS       sequence-384597-        5813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for GFP tagged NHERF1. Dual PDZ1/2 
            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 PDZ1/2*
FEATURES             Location/Qualifiers
     source          1..5813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        162..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      708..717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(714..716,717..719,720..1430)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(714..716,717..719,720..1430)
                     /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(759..780)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1020..1039)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1367..1388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2702..2823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2739..2758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2793..2812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2830..3285)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2967..2988)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3179..3198
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3312..3416
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3418..3775
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3418..3609
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3441..3461)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3626..3761
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3810..4604
                     /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             3810..4604
                     /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(3864..3883)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4474..4493
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4792..4811)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4836..4883
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5212..5800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5701..5720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
       61 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc accatggtga
      721 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      781 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      841 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      901 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      961 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1021 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1081 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1141 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1201 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1261 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1321 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1381 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggccgga
     1441 ctcagatcct gatgagcgcg gacgcagcgg ccggggcgcc cctgccccgg ctctgctgcc
     1501 tggagaaggg tccgaacggc tacgccgccc acctgcacgg ggagaagggc aagttgggcc
     1561 agtacatccg gctggtggag cccggctcgc cggccgagaa ggcggggctg ctggcggggg
     1621 accggctggt ggaggtgaac ggggagaacg tggaaaagga gacccaccag caggtggtga
     1681 gccgcatccg cgccgcactc aacgccgtgc gcctgctggt ggtcgacccc gagacggacg
     1741 agcagctgca gaagctcggc gtccaggtcc gagaggagct gctgcgcgcc caggaagcgc
     1801 cggggcaggc cgagccgccg gccgccgccg aggtgcaggg ggctggcaac gaaaatgagc
     1861 ctcgcgaggc cgacaagagc cacccggagc agcgcgagct tcggcctcgg ctctgtacca
     1921 tgaagaaggg ccccagtggc tatgccgcca acctgcacag cgacaagtcc aagccaggcc
     1981 agttcatccg gtcagtggac ccagactccc cggctgaggc ttcagggctc cgggcccagg
     2041 atcgcattgt ggaggtgaac ggggtctgca tggaggggaa gcagcatggg gacgtggtgt
     2101 ccgccatcag ggctggcggg gacgagacca agctgctggt ggtggacagg gaaactgacg
     2161 agttcttcaa gaaatgcaga gtgatcccat ctcaggagca cctgaatggt cccctgcctg
     2221 tgcccttcac caatggggag atacagaagg agaacagtcg tgaagccctg gcagaggcag
     2281 ccttggagag ccccaggcca gccctggtga gatccgcctc cagtgacacc agcgaggagc
     2341 tgaattccca agacagcccc ccaaaacagg actccacagc gccctcgtct acctcctcct
     2401 ccgaccccat cctagacttc aacatctccc tggccatggc caaagagagg gcccaccaga
     2461 aacgcagcag caaacgggcc ccgcagatgg actggagcaa gaaaaacgaa ctcttcagca
     2521 acctctgagc cgggtcgagg ggaattctgc agtcgacggt accgcgggcc cgggatccac
     2581 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2641 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2701 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2761 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2821 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2881 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2941 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3001 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3061 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3121 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3181 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3241 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3301 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3361 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3421 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3481 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3601 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3661 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3721 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3781 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3841 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3901 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3961 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4021 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4081 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4141 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4201 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4261 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4321 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4381 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4441 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4501 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4561 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4621 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4681 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4741 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4801 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4861 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4921 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4981 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5041 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5101 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5161 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5221 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5281 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5341 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5401 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5461 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5521 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5581 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5641 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5701 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5761 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acg
//
