LOCUS       sequence-158999-        7429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length human alpha ENaC with Flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7429)
  AUTHORS   Raikwar NS, Thomas CP
  TITLE     Nedd4-2 isoforms ubiquitinate individual epithelial sodium channel 
            subunits and reduce surface expression and function of the 
            epithelial sodium channel.
  JOURNAL   Am J Physiol Renal Physiol. 2008 May;294(5):F1157-65. doi: 
            10.1152/ajprenal.00339.2007. Epub 2008 Mar 5.
  PUBMED    18322022
REFERENCE   2  (bases 1 to 7429)
  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: 
            "10.1152/ajprenal.00339.2007"; journalName: "Am J Physiol Renal 
            Physiol"; date: "2008-05"; volume: "294"; issue: "5"; pages: 
            "F1157-65"
COMMENT     Sequence Label: h-alpha-ENaC
FEATURES             Location/Qualifiers
     source          1..7429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2927..2950
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2981..2999)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2982..2999)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3019..3036)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3025..3249
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3295..3723
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3382..3401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3592..3613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3732..3752)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3737..4067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3918..4053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3980..3999
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4134..4928
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4188..4207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4798..4817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5102..5223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5139..5158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5193..5212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5272..5288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5272..5288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5285..5307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5296..5312
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5320..5326,5327..5344,5345..5350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5365..5386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5503..5520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5674..6262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5754..5773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6433..7224,7225..7293))
                     /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     7056..7075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7294..7398)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcatggagg ggaacaagct
      961 ggaggagcag gactctagcc ctccacagtc cactccaggg ctcatgaagg ggaacaagcg
     1021 tgaggagcag gggctgggcc ccgaacctgc ggcgccccag cagcccacgg cggaggagga
     1081 ggccctgatc gagttccacc gctcctaccg agagctcttc gagttcttct gcaacaacac
     1141 caccatccac ggcgccatcc gcctggtgtg ctcccagcac aaccgcatga agacggcctt
     1201 ctgggcagtg ctgtggctct gcacctttgg catgatgtac tggcaattcg gcctgctttt
     1261 cggagagtac ttcagctacc ccgtcagcct caacatcaac ctcaactcgg acaagctcgt
     1321 cttccccgca gtgaccatct gcaccctcaa tccctacagg tacccggaaa ttaaagagga
     1381 gctggaggag ctggaccgca tcacagagca gacgctcttt gacctgtaca aatacagctc
     1441 cttcaccact ctcgtggccg gctcccgcag ccgtcgcgac ctgcggggga ctctgccgca
     1501 ccccttgcag cgcctgaggg tcccgccccc gcctcacggg gcccgtcgag cccgtagcgt
     1561 ggcctccagc ttgcgggaca acaaccccca ggtggactgg aaggactgga agatcggctt
     1621 ccagctgtgc aaccagaaca aatcggactg cttctaccag acatactcat caggggtgga
     1681 tgcggtgagg gagtggtacc gcttccacta catcaacatc ctgtcgaggc tgccagagac
     1741 tctgccatcc ctggaggagg acacgctggg caacttcatc ttcgcctgcc gcttcaacca
     1801 ggtctcctgc aaccaggcga attactctca cttccaccac ccgatgtatg gaaactgcta
     1861 tactttcaat gacaagaaca actccaacct ctggatgtct tccatgcctg gaattaacaa
     1921 cggtctgtcc ctgatgctgc gcgcagagca gaatgacttc attcccctcc tgtccacagt
     1981 gactggggcc cgggtaatgg tgcacgggca ggatgaacct gcctttatgg atgatggtgg
     2041 ctttaacttg cggcctggcg tggagacctc catcagcatg aggaaggaaa ccctggacag
     2101 acttgggggc gattatggcg actgcaccaa gaatggcagt gatgttcctg ttgagaacct
     2161 ttacccttcc aagtacacac agcaggtgtg tattcactcc tgcttccagg agagcatgat
     2221 caaggagtgt ggctgtgcct acatcttcta tccgcggccc cagaacgtgg agtactgtga
     2281 ctacagaaag cacagttcct gggggtactg ctactataag ctccaggttg acttctcctc
     2341 agaccacctg ggctgtttca ccaagtgccg gaagccatgc agcgtgacca gctaccagct
     2401 ctctgctggt tactcacgat ggccctcggt gacatcccag gaatgggtct tccagatgct
     2461 atcgcgacag aacaattaca ccgtcaacaa caagagaaat ggagtggcca aagtcaacat
     2521 cttcttcaag gagctgaact acaaaaccaa ttctgagtct ccctctgtca cgatggtcac
     2581 cctcctgtcc aacctgggca gccagtggag cctgtggttc ggctcctcgg tgttgtctgt
     2641 ggtggagatg gctgagctcg tctttgacct gctggtcatc atgttcctca tgctgctccg
     2701 aaggttccga agccgatact ggtctccagg ccgagggggc aggggtgctc aggaggtagc
     2761 ctccaccctg gcatcctccc ctccttccca cttctgcccc caccccatgt ctctgtcctt
     2821 gtcccagcca ggccctgctc cctctccagc cttgacagcc cctccccctg cctatgccac
     2881 cctgggcccc cgcccatctc cagggggctc tgcaggggcc agttccgact acaaggacga
     2941 cgatgacaag tgactcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     3001 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3061 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3121 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3181 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3241 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3301 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3361 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3421 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     3481 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3541 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3601 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3661 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3721 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4081 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4141 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4201 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4261 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4321 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4381 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4441 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4501 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4561 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4621 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4681 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4741 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4801 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4861 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4921 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4981 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5041 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5101 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5161 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5221 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5281 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5341 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5401 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5461 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5521 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5581 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     5881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
