LOCUS       sequence-271716-        7831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Occludin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7831)
  AUTHORS   Balda MS, Whitney JA, Flores C, Gonzalez S, Cereijido M, Matter K
  TITLE     Functional dissociation of paracellular permeability and 
            transepithelial electrical resistance and disruption of the 
            apical-basolateral intramembrane diffusion barrier by expression of 
            a mutant tight junction membrane protein.
  JOURNAL   J Cell Biol. 1996 Aug;134(4):1031-49. doi: 10.1083/jcb.134.4.1031.
  PUBMED    8769425
REFERENCE   2  (bases 1 to 7831)
  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: "10.1083/jcb.134.4";
            journalName: "J Cell Biol"; date: "1996-08"; volume: "134"; issue: 
            "4"; pages: "1031-49"
COMMENT     Sequence Label: pCB6-Occludin
FEATURES             Location/Qualifiers
     source          1..7831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2273..2895
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2472..2491)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2919..2939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2937..3234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3288..4082
                     /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(3342..3361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3952..3971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4734..4868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4771..4790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4825..4844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4873..4892)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4874..4892)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4906..4922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4906..4922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4919..4941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4930..4946
                     /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(4954..4960,4961..4978,4979..4984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4999..5020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5137..5154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5308..5896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5388..5407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6067..6858,6859..6927))
                     /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     6690..6709
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6928..7032)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7059..7514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7146..7165)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7356..7377
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7640..7662
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7654..7671
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7655..7671
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaatt cagatcggaa ttccgatctg gtaccgggcc ccccctcgag cggccgccag
      661 tgtgatggat atctgcagaa ttcggcttgc caccatgttc agcaagaagt cctacgacgg
      721 cccccccgcg gggtacggcc cccccacggg gtacggcgcc cccacggctg attacggcta
      781 cgggtctccg ccgccgggct cctactacgt ggacgacgct ccgcagctct tctacaagtg
      841 gacgtcgccg cccggcgcgg tgcgggggct gcaggcgggg gtcctcgtgc tgtgcatcgc
      901 catcttcgcc tgcgtcgctt ccacgctcgc ctgggattac ggctacggcc tggggggggc
      961 gtacggcacc gggctggggg ggttctacgg ctccaactac tacggcagcg ggctgagcta
     1021 cagctacggc tacgggggct actacggagg ggtgaaccag cgcacggcca acggcttcat
     1081 gatcgccatg gccgtgctgt gcttcctggc ccagctgggg ctgctggtgg ccgcgctcag
     1141 caaatccggg gccacgcgct cgcggcgctt ctacctggcc gtgctggtgc tgagcgccgt
     1201 gctggccttc gtcatgctca tcgcctccat cgtctacatc atgggcgtca acccgcaggc
     1261 gcagatgtcc agcggttact actacagccc cttgttggcc atgtgcagcc aggcctacgg
     1321 cagcacctac ctcaaccagt acatctacca ctactgcacc gtggaccccc aggaggctgt
     1381 ggctgctgtc tgtgggttcc tcatcgtcat cctgctctgc ctcatctgct tcttcgccca
     1441 gaagacgcgc agtaagatct ggcgctacgg caaagccaac atctactggg accgcgcgcc
     1501 cgtggtgcag gaggggcctg acgtggagga gtgggtgaag aacgtggcgg atggggccag
     1561 cgtgcaggac gagacggcca cgcttgccta ctcggagaag cccaccagcc ctgtcgccgc
     1621 ccccccctac agctacgtgc ccccccccag cgctgggtac tacccctcgg gcacctacag
     1681 cagccggggc gaccagccgg accgggccct cagtgccagc cctgtgcatg gggaggagga
     1741 ggaggagaag gggaaggatc agcccagcag accgcccgcc cgccggggcc gccgccgccg
     1801 ccgtaacccc gagttggatg agtcccagta tgagaccgac tacaccacgg ccgtggagtc
     1861 cagtgatgag cgggaccagg agcagtgggc cagtctgtac ccccccatca cgtcggacgg
     1921 cgcccgccag cgctacaagc aggagttcga caccgacctg aagcgctaca agcagctctg
     1981 tgctgagatg gacagcatca acgaccgcct caatcagctc agccgacggc tcgacagcat
     2041 caccgaggac agccctcaat accaggatgt ggcagaggag tacaatcagc tcaaagacct
     2101 gaagcggagc ccagactacc aaagcaagaa gcaggagagc aaagtgctgc gcaacaagct
     2161 cttccacatc aagcgcatgg tgagcgccta cgacaaggtg cgggggtaaa agccgaattc
     2221 tgcagatatc catcacactg gcggccgctc gagcatgcat ctagaggatc ccgggtggca
     2281 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     2341 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     2401 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     2461 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     2521 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     2581 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     2641 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     2701 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa
     2761 ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc
     2821 ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta
     2881 gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3061 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3121 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3181 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggaat
     3241 tgatcagctt gggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3301 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3361 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3421 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3481 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3541 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3601 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3661 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3721 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3781 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3841 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3901 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3961 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4021 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4081 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccgggct
     4261 cgatcccctc gcgagttggt tcagctgctg cctgaggctg gacgacctcg cggagttcta
     4321 ccggcagtgc aaatccgtcg gcatccagga aaccagcagc ggctatccgc gcatccatgc
     4381 ccccgaactg caggagtggg gaggcacgat ggccgctttg gtcgacccgg acgggacgct
     4441 cctgcgcctg atacagaacg aattgcttgc aggcatctca tgagtgtgtc ttcccgtttt
     4501 ccgcctgagg tcactgcgtg gatggagcgc tggcgcctgc tgcgcgacgg cgagctgctc
     4561 accacccact cgccaagctg gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4621 ctccgccgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4681 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4741 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4801 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4861 tctggatcaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc
     4921 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4981 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5041 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5101 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5161 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5221 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5941 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6001 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6061 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6121 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6181 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6241 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6301 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6361 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6421 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6481 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6541 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6601 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6661 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6721 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6781 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6841 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6901 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6961 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7021 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     7081 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     7141 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7201 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7261 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7321 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7381 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7441 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7501 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7561 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7621 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
     7681 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
     7741 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     7801 attggctcat gtccaatatg accgccatgt t
//
