LOCUS       sequence-271724-        7805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HAOccludin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7805)
  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 7805)
  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-HAOccludin
FEATURES             Location/Qualifiers
     source          1..7805
                     /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    2247..2869
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2446..2465)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2893..2913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2911..3208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3262..4056
                     /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(3316..3335)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3926..3945
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4708..4842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4745..4764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4799..4818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4847..4866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4848..4866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4880..4896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4880..4896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4893..4915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4904..4920
                     /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(4928..4934,4935..4952,4953..4958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4973..4994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5111..5128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5282..5870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5362..5381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6041..6832,6833..6901))
                     /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     6664..6683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6902..7006)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7033..7488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7120..7139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7330..7351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7614..7636
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7628..7645
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7629..7645
                     /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 gtaccatgca ggacctgcca ggcaacgaca
      661 acagcaccgc cggccttaag aagtcctacg acggcccccc cgcggggtac ggccccccca
      721 cggggtacgg cgcccccacg gctgattacg gctacgggtc tccgccgccg ggctcctact
      781 acgtggacga cgctccgcag ctcttctaca agtggacgtc gccgcccggc gcggtgcggg
      841 ggctgcaggc gggggtcctc gtgctgtgca tcgccatctt cgcctgcgtc gcttccacgc
      901 tcgcctggga ttacggctac ggcctggggg gggcgtacgg caccgggctg ggggggttct
      961 acggctccaa ctactacggc agcgggctga gctacagcta cggctacggg ggctactacg
     1021 gaggggtgaa ccagcgcacg gccaacggct tcatgatcgc catggccgtg ctgtgcttcc
     1081 tggcccagct ggggctgctg gtggccgcgc tcagcaaatc cggggccacg cgctcgcggc
     1141 gcttctacct ggccgtgctg gtgctgagcg ccgtgctggc cttcgtcatg ctcatcgcct
     1201 ccatcgtcta catcatgggc gtcaacccgc aggcgcagat gtccagcggt tactactaca
     1261 gccccttgtt ggccatgtgc agccaggcct acggcagcac ctacctcaac cagtacatct
     1321 accactactg caccgtggac ccccaggagg ctgtggctgc tgtctgtggg ttcctcatcg
     1381 tcatcctgct ctgcctcatc tgcttcttcg cccagaagac gcgcagtaag atctggcgct
     1441 acggcaaagc caacatctac tgggaccgcg cgcccgtggt gcaggagggg cctgacgtgg
     1501 aggagtgggt gaagaacgtg gcggatgggg ccagcgtgca ggacgagacg gccacgcttg
     1561 cctactcgga gaagcccacc agccctgtcg ccgccccccc ctacagctac gtgccccccc
     1621 ccagcgctgg gtactacccc tcgggcacct acagcagccg gggcgaccag ccggaccggg
     1681 ccctcagtgc cagccctgtg catggggagg aggaggagga gaaggggaag gatcagccca
     1741 gcagaccgcc cgcccgccgg ggccgccgcc gccgccgtaa ccccgagttg gatgagtccc
     1801 agtatgagac cgactacacc acggccgtgg agtccagtga tgagcgggac caggagcagt
     1861 gggccagtct gtaccccccc atcacgtcgg acggcgcccg ccagcgctac aagcaggagt
     1921 tcgacaccga cctgaagcgc tacaagcagc tctgtgctga gatggacagc atcaacgacc
     1981 gcctcaatca gctcagccga cggctcgaca gcatcaccga ggacagccct caataccagg
     2041 atgtggcaga ggagtacaat cagctcaaag acctgaagcg gagcccagac taccaaagca
     2101 agaagcagga gagcaaagtg ctgcgcaaca agctcttcca catcaagcgc atggtgagcg
     2161 cctacgacaa ggtgcggggg taaaagccga attctgcaga tatccatcac actggcggcc
     2221 gctcgagcat gcatctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc
     2281 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     2341 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     2401 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     2461 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     2521 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     2581 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     2641 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     2701 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc
     2761 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg
     2821 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc
     2881 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3181 tccagaagta gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag
     3241 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3301 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3361 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3421 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3481 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3541 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3601 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3661 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3721 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3781 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3841 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3901 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3961 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4021 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4081 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4141 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4201 atctcatgct ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct
     4261 gctgcctgag gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc
     4321 aggaaaccag cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca
     4381 cgatggccgc tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc
     4441 ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga
     4501 gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg
     4561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc
     4621 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4681 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4801 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc
     4861 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4921 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4981 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5041 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6181 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7021 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     7081 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     7141 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     7201 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     7261 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7321 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     7381 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     7441 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     7501 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7561 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7621 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     7681 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     7741 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     7801 atgtt
//
