LOCUS       sequence-171580-        6197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length Occludin with mutation of PPPY domain at AA9-12 to PAAY.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  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 6197)
  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: OccludinMut_Flag
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(116..134)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(148..164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(148..164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(161..183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    172..188
                     /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(196..202,203..220,221..226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    241..262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(379..396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(550..1138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(630..649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1309..2100,2101..2169))
                     /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     1932..1951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2170..2274)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2301..2756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2388..2407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2598..2619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2882..2904
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2896..2913
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2897..2913
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3074..3453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3454..3657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3608..3628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3693..3716
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5493..5512)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5940..5960)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(5945..6197,1..77)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6125..6197,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
       61 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat
      121 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
      181 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      241 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      301 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      361 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      421 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      481 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      961 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2281 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2701 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat
     2761 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2821 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2881 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac attgaatcaa
     2941 tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg
     3001 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
     3061 tgaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     3121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     3241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     3301 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
     3361 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
     3421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat
     3481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     3541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
     3601 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     3661 ttagtgaacc gtcagaatta ggccttacca tggactacaa agacgatgac gacaagctca
     3721 aagcttcatc caggcctctt gaaagtccag ctgcttacag gcctgatgaa ttcaaaccga
     3781 atcattatgc accaagcaat gacatatatg gtggagagat gcatgttcga ccaatgctct
     3841 ctcagccagc ctactctttt tacccagaag atgaaactct tcacttctac aaatggacct
     3901 ctcctccagg agtgattcgg atcctgtcta tgctcattat tgtgatgtgc attgccatct
     3961 ttgcctgtgt ggcctccacg cttgcctggg acagaggcta cggaacttcc cttttaggag
     4021 gtagtgtagg ctacccttat ggaggaagtg gctttggtag ctacggaagt ggctatggct
     4081 atggctatgg ttatggctat ggctacggag gctatacaga cccaagagca gcaaagggct
     4141 tcatgttggc catggctgcc ttttgtttca ttgccgcgtt ggtgatcttt gttaccagtg
     4201 ttataagatc tgaaatgtcc agaacaagaa gatactactt aagtgtgata atagtgagtg
     4261 ctatcctggg catcatggtg tttattgcca caattgtcta tataatggga gtgaacccaa
     4321 ctgctcagtc ttctggatct ctatatggtt cacaaatata tgccctctgc aaccaatttt
     4381 atacacctgc agctactgga ctctacgtgg atcagtattt gtatcactac tgtgttgtgg
     4441 atccccagga ggccattgcc attgtactgg ggttcatgat tattgtggct tttgctttaa
     4501 taattttctt tgctgtgaaa actcgaagaa agatggacag gtatgacaag tccaatattt
     4561 tgtgggacaa ggaacacatt tatgatgagc agccccccaa tgtcgaggag tgggttaaaa
     4621 atgtgtctgc aggcacacag gacgtgcctt cacccccatc tgactatgtg gaaagagttg
     4681 acagtcccat ggcatactct tccaatggca aagtgaatga caagcggttt tatccagagt
     4741 cttcctataa atccacgccg gttcctgaag tggttcagga gcttccatta acttcgcctg
     4801 tggatgactt caggccgcct cgttacagca gcggtggtaa ctttgagaca ccttcaaaaa
     4861 gagcacctgc aaagggaaga gcaggaaggt caaagagaac agagcaagat cactatgaga
     4921 cagactacac aactggcggc gagtcctgtg atgagctgga ggaggactgg atcagggaat
     4981 atccacctat cacttcagat caacaaagac aactgtacaa gaggaatttt gacactggcc
     5041 tacaggaata caagagctta caatcagaac ttgatgagat caataaagaa ctctcccgtt
     5101 tggataaaga attggatgac tatagagaag aaagtgaaga gtacatggct gctgctgatg
     5161 aatacaatag actgaagcaa gtgaagggat ctgcagatta caaaagtaag aagaatcatt
     5221 gcaagcagtt aaagagcaaa ttgtcacaca tcaagaagat ggttggagac tatgatagac
     5281 agaaaacata gcccggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     5341 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     5401 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     5461 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     5521 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     5581 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     5641 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     5701 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     5761 ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag
     5821 gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc
     5881 atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag cttggctgtg
     5941 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6001 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     6061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     6121 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6181 tttttttatt tatgcag
//
