LOCUS       sequence-158992-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nedd4L-deltaC2 with His Xpress tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  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 7897)
  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 delta C2
FEATURES             Location/Qualifiers
     source          1..7897
                     /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(3904..3921)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3910..4134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4180..4608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4267..4286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4477..4498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4617..4637)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4622..4952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4803..4938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4865..4884
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5000..5047
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5066..5440
                     /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    5570..5691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5607..5626)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5661..5680
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5740..5756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5740..5756)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5753..5775)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5764..5780
                     /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(5788..5794,5795..5812,5813..5818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5833..5854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5971..5988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6142..6730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6222..6241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6901..7692,7693..7761))
                     /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     7524..7543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7762..7866)
                     /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 aggtaccagg atccatggag cgaccctata
     1201 catttaagga ctttctcctc agaccaagaa gtcataagtc tcgagttaag ggatttttgc
     1261 gattgaaaat ggcctatatg ccaaaaaatg gaggtcaaga tgaagaaaac agtgaccaga
     1321 gggatgacat ggagcatgga tgggaagttg ttgactcaaa tgactcggct tctcagcacc
     1381 aagaggaact tcctcctcct cctctgcctc ccgggtggga agaaaaagtg gacaatttag
     1441 gccgaactta ctatgtcaac cacaacaacc ggaccactca gtggcacaga ccaagcctga
     1501 tggacgtgtc ctcggagtcg gacaataaca tcagacagat caaccaggag gcagcacacc
     1561 ggcgcttccg ctcccgcagg cacatcagcg aagacttgga gcccgagccc tcggagggcg
     1621 gggatgtccc cgagccttgg gagaccattt cagaggaagt gaatatcgct ggagactctc
     1681 tcggtctggc tctgccccca ccaccggcct ccccaggatc tcggaccagc cctcaggagc
     1741 tgtcagagga actaagcaga aggcttcaga tcactccaga ctccaatggg gaacagttca
     1801 gctctttgat tcaaagagaa ccctcctcaa ggttgaggtc atgcagtgtc accgacgcag
     1861 ttgcagaaca gggccatcta ccaccgccca gtgccccagc tgggagagcg cgttcatcaa
     1921 ctgtcacggg tggtgaggaa ccaacgccat cagtggccta tgtacatacc acgccgggtc
     1981 tgccttcagg ctgggaagaa agaaaagatg ctaaggggcg cacatactat gtcaatcata
     2041 acaatcgaac cacaacttgg actcgaccta tcatgcagct tgcagaagat ggtgcgtccg
     2101 gatcagccac aaacagtaac aaccatctaa tcgagcctca gatccgccgg cctcgtagcc
     2161 tcagctcgcc aacagtaact ttatctgccc cgctggaggg tgccaaggac tcacccgtac
     2221 gtcgggctgt gaaagacacc ctttccaacc cacagtcccc acagccatca ccttacaact
     2281 cccccaaacc acaacacaaa gtcacacaga gcttcttgcc acccggctgg gaaatgagga
     2341 tagcgccaaa cggccggccc ttcttcattg atcataacac aaagactaca acctgggaag
     2401 atccacgttt gaaatttcca gtacatatgc ggtcaaagac atctttaaac cccaatgacc
     2461 ttggccccct tcctcctggc tgggaagaaa gaattcactt ggatggccga acgttttata
     2521 ttgatcataa tagcaaaatt actcagtggg aagacccaag actgcagaac ccagctatta
     2581 ctggtccggc tgtcccttac tccagagaat ttaagcagaa atatgactac ttcaggaaga
     2641 aattaaagaa acctgctgat atccccaata ggtttgaaat gaaacttcac agaaataaca
     2701 tatttgaaga gtcctatcgg agaattatgt ccgtgaaaag accagatgtc ctaaaagcta
     2761 gactgtggat tgagtttgaa tcagagaaag gtcttgacta tgggggtgtg gccagagaat
     2821 ggttcttctt actgtccaaa gagatgttca acccctacta cggcctcttt gagtactctg
     2881 ccacggacaa ctacaccctt cagatcaacc ctaattcagg cctctgtaat gaggatcatt
     2941 tgtcctactt cacttttatt ggaagagttg ctggtctggc cgtatttcat gggaagctct
     3001 tagatggttt cttcattaga ccattttaca agatgatgtt gggaaagcag ataaccctga
     3061 atgacatgga atctgtggat agtgaatatt acaactcttt gaaatggatc ctggagaatg
     3121 accctactga gctggacctc atgttctgca tagacgaaga aaactttgga cagacatatc
     3181 aagtggattt gaagcccaat gggtcagaaa taatggtcac aaatgaaaac aaaagggaat
     3241 atatcgactt agtcatccag tggagatttg tgaacagggt ccagaagcag atgaacgcat
     3301 tcttggaggg attcacagaa ctacttccta ttgatttgat taaaattttt gatgaaaatg
     3361 agctggagtt gctcatgtgc ggcctcggtg atgtggatgt gaatgactgg agacagcatt
     3421 ctatttacaa gaacggctac tgcccaaacc accccgtcat tcagtggttc tggaaggctg
     3481 tgctactcat ggacgccgaa aagcgtatcc ggttactgca gtttgtcaca gggacatcgc
     3541 gagtacctat gaatggattt gccgaacttt atggttccaa tggtcctcag ctgtttacaa
     3601 tagagcaatg gggcagtcct gagaaactgc ccagagctca cacatgcttt aatcgccttg
     3661 acttacctcc atatgaaacc tttgaagatt tacgagagaa acttctcatg gccgtggaaa
     3721 atgctcaagg atttgaaggg gtggattaag caccctgtac ctcgggggtg gttgttcttc
     3781 aagcaagttc tgcttgcact tttgcatttg cctaacagac ttttgcagag gcgatggaag
     3841 ggcgaattcc agcacactgg cggccgttac tagctagagg gcccgtttaa acccgctgat
     3901 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3961 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4021 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4081 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4141 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4201 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4261 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4321 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4381 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4441 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4501 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     4561 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4621 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     4681 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4861 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4921 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4981 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     5041 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     5101 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     5161 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     5221 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     5281 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     5341 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     5401 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     5461 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5521 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5581 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5641 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5701 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5761 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5821 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5881 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5941 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6001 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6061 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6301 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6541 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6721 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6841 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6901 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6961 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7021 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7081 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7141 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7201 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7261 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7321 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7381 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7441 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7501 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7561 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7621 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7681 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7741 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7801 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7861 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
