LOCUS       sequence-158994-        8259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length Nedd4L with His Xpress tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8259)
  AUTHORS   Itani OA, Stokes JB, Thomas CP
  TITLE     Nedd4-2 isoforms differentially associate with ENaC and regulate its
            activity.
  JOURNAL   Am J Physiol Renal Physiol. 2005 Aug;289(2):F334-46. Epub 2005 Apr 
            5.
  PUBMED    15814530
REFERENCE   2  (bases 1 to 8259)
  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 Renal Physiol."; date: "2005-08"; volume: "289(2)"; pages: 
            "F334-46. Epub 2005 Apr 5"
COMMENT     Sequence Label: hNedd4-2
FEATURES             Location/Qualifiers
     source          1..8259
                     /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"
     misc_feature    917..1079
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             1092..1109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1113..1145
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1149..1157,1158..1172)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(4266..4283)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4272..4496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4542..4970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4629..4648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4839..4860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4979..4999)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4984..5314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5165..5300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5227..5246
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5362..5409
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5428..5802
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5932..6053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5969..5988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6023..6042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6102..6118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6102..6118)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6115..6137)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6126..6142
                     /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(6150..6156,6157..6174,6175..6180))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6195..6216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6333..6350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6504..7092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6584..6603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7263..8054,8055..8123))
                     /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     7886..7905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8124..8228)
                     /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 gctggctagc
      901 gtttaaactt aagcttagcg cagaggcttg gggcagccga gcggcagcca ggccccggcc
      961 cgggcctcgg ttccagaagg gagaggagcc cgccaaggcg cgcaagagag cgggctgcct
     1021 cgcagtccga gccggagagg gagcgcgagc cgcgccggcc ccggacggcc tccgaaacca
     1081 tggggggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
     1141 tgggtcggga tctgtacgac gatgacgata aggtacgcgc cggctccatg gcgaccgggc
     1201 tcggggagcc ggtctatgga ctttccgaag acgagggaga gtcccgtatt ctcagagtaa
     1261 aagttgtttc tggaattgat ctcgccaaaa aggacatctt tggagccagt gatccgtatg
     1321 tgaaactttc attgtacgta gcggatgaga atagagaact tgctttggtc cagacaaaaa
     1381 caattaaaaa gacactgaac ccaaaatgga atgaagaatt ttatttcagg gtaaacccat
     1441 ctaatcacag actcctattt gaagtatttg acgaaaatag actgacacga gacgacttcc
     1501 tgggccaggt ggacgtgccc cttagtcacc ttccgacaga agatccaacc atggagcgac
     1561 cctatacatt taaggacttt ctcctcagac caagaagtca taagtctcga gttaagggat
     1621 ttttgcgatt gaaaatggcc tatatgccaa aaaatggagg tcaagatgaa gaaaacagtg
     1681 accagaggga tgacatggag catggatggg aagttgttga ctcaaatgac tcggcttctc
     1741 agcaccaaga ggaacttcct cctcctcctc tgcctcccgg gtgggaagaa aaagtggaca
     1801 atttaggccg aacttactat gtcaaccaca acaaccggac cactcagtgg cacagaccaa
     1861 gcctgatgga cgtgtcctcg gagtcggaca ataacatcag acagatcaac caggaggcag
     1921 cacaccggcg cttccgctcc cgcaggcaca tcagcgaaga cttggagccc gagccctcgg
     1981 agggcgggga tgtccccgag ccttgggaga ccatttcaga ggaagtgaat atcgctggag
     2041 actctctcgg tctggctctg cccccaccac cggcctcccc aggatctcgg accagccctc
     2101 aggagctgtc agaggaacta agcagaaggc ttcagatcac tccagactcc aatggggaac
     2161 agttcagctc tttgattcaa agagaaccct cctcaaggtt gaggtcatgc agtgtcaccg
     2221 acgcagttgc agaacagggc catctaccac cgcccagtgc cccagctggg agagcgcgtt
     2281 catcaactgt cacgggtggt gaggaaccaa cgccatcagt ggcctatgta cataccacgc
     2341 cgggtctgcc ttcaggctgg gaagaaagaa aagatgctaa ggggcgcaca tactatgtca
     2401 atcataacaa tcgaaccaca acttggactc gacctatcat gcagcttgca gaagatggtg
     2461 cgtccggatc agccacaaac agtaacaacc atctaatcga gcctcagatc cgccggcctc
     2521 gtagcctcag ctcgccaaca gtaactttat ctgccccgct ggagggtgcc aaggactcac
     2581 ccgtacgtcg ggctgtgaaa gacacccttt ccaacccaca gtccccacag ccatcacctt
     2641 acaactcccc caaaccacaa cacaaagtca cacagagctt cttgccaccc ggctgggaaa
     2701 tgaggatagc gccaaacggc cggcccttct tcattgatca taacacaaag actacaacct
     2761 gggaagatcc acgtttgaaa tttccagtac atatgcggtc aaagacatct ttaaacccca
     2821 atgaccttgg cccccttcct cctggctggg aagaaagaat tcacttggat ggccgaacgt
     2881 tttatattga tcataatagc aaaattactc agtgggaaga cccaagactg cagaacccag
     2941 ctattactgg tccggctgtc ccttactcca gagaatttaa gcagaaatat gactacttca
     3001 ggaagaaatt aaagaaacct gctgatatcc ccaataggtt tgaaatgaaa cttcacagaa
     3061 ataacatatt tgaagagtcc tatcggagaa ttatgtccgt gaaaagacca gatgtcctaa
     3121 aagctagact gtggattgag tttgaatcag agaaaggtct tgactatggg ggtgtggcca
     3181 gagaatggtt cttcttactg tccaaagaga tgttcaaccc ctactacggc ctctttgagt
     3241 actctgccac ggacaactac acccttcaga tcaaccctaa ttcaggcctc tgtaatgagg
     3301 atcatttgtc ctacttcact tttattggaa gagttgctgg tctggccgta tttcatggga
     3361 agctcttaga tggtttcttc attagaccat tttacaagat gatgttggga aagcagataa
     3421 ccctgaatga catggaatct gtggatagtg aatattacaa ctctttgaaa tggatcctgg
     3481 agaatgaccc tactgagctg gacctcatgt tctgcataga cgaagaaaac tttggacaga
     3541 catatcaagt ggatttgaag cccaatgggt cagaaataat ggtcacaaat gaaaacaaaa
     3601 gggaatatat cgacttagtc atccagtgga gatttgtgaa cagggtccag aagcagatga
     3661 acgccttctt ggagggattc acagaactac ttcctattga tttgattaaa atttttgacg
     3721 aaaatgagct ggagttgctc atgtgcggcc tcggtgatgt ggatgtgaat gactggagac
     3781 agcattctat ttacaagaac ggctactgcc caaaccaccc cgtcattcag tggttctgga
     3841 aggctgtgct actcatggac gccgaaaagc gtatccggtt actgcagttt gtcacaggga
     3901 catcgcgagt acctatgaat ggatttgccg aactttatgg ttccaatggt cctcagctgt
     3961 ttacaataga gcaatggggc agtcctgaga aactgcccag agctcacaca tgctttaatc
     4021 gccttgactt acctccatat gaaacctttg aagatttacg agagaaactt ctcatggccg
     4081 tggaaaatgc tcaaggattt gaaggggtgg attaagcacc ctgtgcctcg ggggtggttg
     4141 ttcttcaagc aagttctgct tgcacttttg catttgccta acagactttt gcagaggcga
     4201 tggaagggcg aattccagca cactggcggc cgttactaga gggcccgttt aaacccgctg
     4261 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4321 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4381 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4441 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4501 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4561 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4621 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4681 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4741 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4801 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4861 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4921 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4981 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     5041 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     5101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5161 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5221 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5281 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     5341 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     5401 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     5461 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     5521 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     5581 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     5641 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     5701 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     5761 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     5821 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5881 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5941 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6001 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6061 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6121 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6181 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6241 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6301 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6361 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6421 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6661 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6901 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7261 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7321 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7381 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7441 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7501 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7561 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7621 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7681 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7741 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7801 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7861 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7921 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7981 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8041 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8221 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
