LOCUS       sequence-156471-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NHERF2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  AUTHORS   Lau AG, Hall RA
  TITLE     Oligomerization of NHERF-1 and NHERF-2 PDZ domains: differential 
            regulation by association with receptor carboxyl-termini and by 
            phosphorylation.
  JOURNAL   Biochemistry. 2001 Jul 24;40(29):8572-80.
  PUBMED    11456497
REFERENCE   2  (bases 1 to 5702)
  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"; journalName: 
            "Biochemistry."; date: "2001-07-24"; volume: "40(29)"; pages: 
            "8572-80"
COMMENT     Sequence Label: pBK-CMV-hNHERF2
FEATURES             Location/Qualifiers
     source          1..5702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             618..644
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      651..660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2093..2112)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2094..2112)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2119..2136)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2119..2135)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2128..2150)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2497..2618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2588..2607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2625..3080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2762..2783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2974..2993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3107..3211
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3213..3570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3213..3404
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3236..3256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3421..3556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3605..4399
                     /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             3605..4399
                     /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(3659..3678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4269..4288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4587..4606)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4631..4678
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5007..5595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5496..5515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agtggatcgc caccatgtat ccatatgacg ttccagacta tgcagaattc gccgccatgg
      661 ccgcgccgga gccgctgcgg ccgcgcctgt gccgcttggt gcgcggagag cagggctacg
      721 gcttccacct gcacggcgag aagggccgcc gcgggcagtt catccggcgc gtggaacccg
      781 gttcccccgc cgaggccgcc gcgctgcgcg ctggggaccg cctggtcgag gtcaacggcg
      841 tcaacgtgga gggcgagacg caccaccagg tggtgcaaag gatcaaggct gtggaggggc
      901 agactcggct gctggtggtg gaccaggaga cagatgagga gctccgccgg cggcagctga
      961 cctgtaccga ggagatggcc cagcgagggc tcccacccgc ccacgacccc tgggagccga
     1021 agccagactg ggcacacacc ggcagccaca gctccgaagc tggcaagaag gatgtcagtg
     1081 ggcccctgag ggagctccgg cctcggctct gccacctgcg aaagggacct cagggctatg
     1141 ggttcaacct gcatagtgac aagtcccggc ccggccagta catccgctct gtggacccgg
     1201 gctcacctgc cgcccgctct ggcctccgcg cccaggaccg gctcattgag gtgaacgggc
     1261 agaatgtgga gggactgcgc catgctgagg tggtggccag catcaaggca cgggaggacg
     1321 aggcccggct gctggtcgtg gaccccgaga cagatgaaca cttcaagcgg cttcgggtca
     1381 cacccaccga ggagcacgtg gaaggtcctc tgccgtcacc cgtcaccaat ggaaccagcc
     1441 ctgcccagct caatggtggc tctgcgtgct cgtcccgaag tgacctgcct ggttccgaca
     1501 aggacactga ggatggcagt gcctggaagc aagatccctt ccaggagagc ggcctccacc
     1561 tgagccccac ggcggccgag gccaaggaga aggctcgagc catgcgagtc aacaagcgcg
     1621 cgccacagat ggactggaac aggaagcgtg aaatcttcag caacttctga gccccttcct
     1681 gcctgtctcg ggaccctggg acccctcccg cacggacctt gggcctcagc ctgccccgag
     1741 ctcccccagc ctcagtggac tggagggtgg tcctgccatt gcccagaaat cagccccagc
     1801 cccggtgagc ccccatcctg cccctgccca ccaggtactg ggggcctgtg gcagcaagat
     1861 agggggagag agacccagag atgtgagaga gagtcagaga cagagacaga gagagagaga
     1921 gagagacaca gagagagaca gagagagagc gagcgagcgc gcggcagccg cggggcgagg
     1981 gcctttgctg cgctgccggg gcctgctgac tgagtcgaga gtacttctag agcggccgcg
     2041 ggcccatcga ttttccaccc gggtggggta ccaggtaagt gtacccaatt cgccctatag
     2101 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2161 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2221 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggagatc
     2281 caatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt
     2341 cacagtccca aggctcattt caggcccctc agtcctcaca gtctgttcat gatcataatc
     2401 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2461 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2521 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2581 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2641 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2701 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2761 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2821 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2881 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2941 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3001 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3061 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3181 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3241 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3421 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3481 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3541 ggcttttttg gaggcctagg cttttgcaaa gatctatcaa gagacaggat gaggatcgtt
     3601 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3661 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3721 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3781 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3841 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3901 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3961 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4021 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4081 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4141 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4201 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4261 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4321 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4381 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4441 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4501 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4561 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4621 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4681 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4741 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4801 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4861 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4921 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4981 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5041 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5101 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5161 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5221 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5281 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5341 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5401 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5461 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5521 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5581 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5641 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5701 gc
//
