LOCUS       sequence-171539-        8258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length NeddL3 with C to S mutation with His Xpress tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8258)
  AUTHORS   Raikwar NS, Vandewalle A, Thomas CP
  TITLE     Nedd4-2 interacts with occludin to inhibit tight junction formation 
            and enhance paracellular conductance in collecting duct epithelia.
  JOURNAL   Am J Physiol Renal Physiol. 2010 Aug;299(2):F436-44. doi: 
            10.1152/ajprenal.00674.2009. Epub 2010 May 26.
  PUBMED    20504882
REFERENCE   2  (bases 1 to 8258)
  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.00674.2009"; journalName: "Am J Physiol Renal 
            Physiol"; date: "2010-08"; volume: "299"; issue: "2"; pages: 
            "F436-44"
COMMENT     Sequence Label: hNedd4-2 CS
FEATURES             Location/Qualifiers
     source          1..8258
                     /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..5313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5164..5299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5226..5245
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5361..5408
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5427..5801
                     /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    5931..6052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5968..5987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6022..6041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6101..6117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6101..6117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6114..6136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6125..6141
                     /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(6149..6155,6156..6173,6174..6179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6194..6215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6332..6349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6503..7091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6583..6602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7262..8053,8054..8122))
                     /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     7885..7904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8123..8227)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 catactacgc
     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 atttttgatg
     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 tcctttaatc
     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 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4741 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4801 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4861 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4921 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4981 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5041 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5101 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5161 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5221 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5281 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5341 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     5401 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     5461 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     5521 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     5581 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     5641 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     5701 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     5761 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     5821 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5881 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6061 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6241 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6661 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6901 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7021 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7081 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7141 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7201 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7261 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7321 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7381 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7441 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7501 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7561 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7621 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7681 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7741 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7801 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7861 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7921 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7981 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8041 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8101 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8161 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8221 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
