LOCUS       sequence-164402-        7625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length human alpha ENaC-2 with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7625)
  AUTHORS   Thomas CP, Auerbach S, Stokes JB, Volk KA
  TITLE     5' heterogeneity in epithelial sodium channel alpha-subunit mRNA 
            leads to distinct NH2-terminal variant proteins.
  JOURNAL   Am J Physiol. 1998 May;274(5 Pt 1):C1312-23.
  PUBMED    9612219
REFERENCE   2  (bases 1 to 7625)
  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: "Am J 
            Physiol."; date: "1998-05"; volume: "274(5 Pt 1)"; pages: "C1312-23"
COMMENT     Sequence Label: h-alpha-ENaC-2
FEATURES             Location/Qualifiers
     source          1..7625
                     /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             3123..3146
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3177..3195)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3178..3195)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3215..3232)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3221..3445
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3491..3919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3578..3597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3788..3809
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3928..3948)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3933..4263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4114..4249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4176..4195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4330..5124
                     /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(4384..4403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4994..5013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5298..5419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5335..5354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5389..5408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5468..5484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5468..5484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5481..5503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5492..5508
                     /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(5516..5522,5523..5540,5541..5546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5561..5582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5699..5716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5870..6458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5950..5969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6629..7420,7421..7489))
                     /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     7252..7271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7490..7594)
                     /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 ttgggcagtc gcagggtggg
      961 acatgggcat ggccaggggc agcctcactc gggttccagg ggtgatggga gagggcactc
     1021 agggcccaga gctcagcctt gaccctgacc cttgctctcc ccaatccact ccggggctca
     1081 tgaaggggaa caagctggag gagcaggacc ctagacctct gcagcccata ccaggtctca
     1141 tggaggggaa caagctggag gagcaggact ctagccctcc acagtccact ccagggctca
     1201 tgaaggggaa caagcgtgag gagcaggggc tgggccccga acctgcggcg ccccagcagc
     1261 ccacggcgga ggaggaggcc ctgatcgagt tccaccgctc ctaccgagag ctcttcgagt
     1321 tcttctgcaa caacaccacc atccacggcg ccatccgcct ggtgtgctcc cagcacaacc
     1381 gcatgaagac ggccttctgg gcagtgctgt ggctctgcac ctttggcatg atgtactggc
     1441 aattcggcct gcttttcgga gagtacttca gctaccccgt cagcctcaac atcaacctca
     1501 actcggacaa gctcgtcttc cccgcagtga ccatctgcac cctcaatccc tacaggtacc
     1561 cggaaattaa agaggagctg gaggagctgg accgcatcac agagcagacg ctctttgacc
     1621 tgtacaaata cagctccttc accactctcg tggccggctc ccgcagccgt cgcgacctgc
     1681 gggggactct gccgcacccc ttgcagcgcc tgagggtccc gcccccgcct cacggggccc
     1741 gtcgagcccg tagcgtggcc tccagcttgc gggacaacaa cccccaggtg gactggaagg
     1801 actggaagat cggcttccag ctgtgcaacc agaacaaatc ggactgcttc taccagacat
     1861 actcatcagg ggtggatgcg gtgagggagt ggtaccgctt ccactacatc aacatcctgt
     1921 cgaggctgcc agagactctg ccatccctgg aggaggacac gctgggcaac ttcatcttcg
     1981 cctgccgctt caaccaggtc tcctgcaacc aggcgaatta ctctcacttc caccacccga
     2041 tgtatggaaa ctgctatact ttcaatgaca agaacaactc caacctctgg atgtcttcca
     2101 tgcctggaat taacaacggt ctgtccctga tgctgcgcgc agagcagaat gacttcattc
     2161 ccctcctgtc cacagtgact ggggcccggg taatggtgca cgggcaggat gaacctgcct
     2221 ttatggatga tggtggcttt aacttgcggc ctggcgtgga gacctccatc agcatgagga
     2281 aggaaaccct ggacagactt gggggcgatt atggcgactg caccaagaat ggcagtgatg
     2341 ttcctgttga gaacctttac ccttcaaagt acacacagca ggtgtgtatt cactcctgct
     2401 tccaggagag catgatcaag gagtgtggct gtgcctacat cttctatccg cggccccaga
     2461 acgtggagta ctgtgactac agaaagcaca gttcctgggg gtactgctac tataagctcc
     2521 aggttgactt ctcctcagac cacctgggct gtttcaccaa gtgccggaag ccatgcagcg
     2581 tgaccagcta ccagctctct gctggttact cacgatggcc ctcggtgaca tcccaggaat
     2641 gggtcttcca gatgctatcg cgacagaaca attacaccgt caacaacaag agaaatggag
     2701 tggccaaagt caacatcttc ttcaaggagc tgaactacaa aaccaattct gagtctccct
     2761 ctgtcacgat ggtcaccctc ctgtccaacc tgggcagcca gtggagcctg tggttcggct
     2821 cctcggtgtt gtctgtggtg gagatggctg agctcgtctt tgacctgctg gtcatcatgt
     2881 tcctcatgct gctccgaagg ttccgaagcc gatactggtc tccaggccga gggggcaggg
     2941 gtgctcagga ggtagcctcc accctggcat cctcccctcc ttcccacttc tgcccccacc
     3001 ccatgtctct gtccttgtcc cagccaggcc ctgctccctc tccagccttg acagcccctc
     3061 cccctgccta tgccaccctg ggcccccgcc catctccagg gggctctgca ggggccagtt
     3121 ccgactacaa ggacgacgat gacaagtgac tcgagcatgc atctagaggg ccctattcta
     3181 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3241 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3301 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3361 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3421 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3481 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3541 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     3661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3781 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3841 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3901 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3961 caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4201 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4261 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4321 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5161 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5221 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5281 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5401 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5461 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5521 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5581 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5641 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5701 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6061 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6301 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6421 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6481 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6541 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6601 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6661 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6721 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6781 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6841 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6901 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6961 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7021 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7081 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7141 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7201 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7261 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7321 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7381 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7441 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7501 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7561 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7621 acgtc
//
