LOCUS       sequence-271738-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of OccludinCT3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  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 7025)
  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-OccludinCT3
FEATURES             Location/Qualifiers
     source          1..7025
                     /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    1467..2089
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1666..1685)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2113..2133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2131..2428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2482..3276
                     /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(2536..2555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3146..3165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3928..4062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3965..3984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4019..4038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4067..4086)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4068..4086)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4100..4116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4100..4116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4113..4135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4124..4140
                     /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(4148..4154,4155..4172,4173..4178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4193..4214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4331..4348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4502..5090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4582..4601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5261..6052,6053..6121))
                     /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     5884..5903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6122..6226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6253..6708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6340..6359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6550..6571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6834..6856
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6848..6865
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6849..6865
                     /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 taatctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc
     1501 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     1561 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     1621 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     1681 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     1741 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     1801 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     1861 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     1921 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc
     1981 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg
     2041 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc
     2101 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2281 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2341 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2401 tccagaagta gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag
     2461 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2521 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2581 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2641 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     2701 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2761 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2821 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2881 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2941 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3001 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3061 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3121 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3181 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3241 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3301 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     3361 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3421 atctcatgct ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct
     3481 gctgcctgag gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc
     3541 aggaaaccag cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca
     3601 cgatggccgc tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc
     3661 ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga
     3721 gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg
     3781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc
     3841 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3901 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3961 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4021 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc
     4081 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4201 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4261 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4921 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6241 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6661 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     6721 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6781 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6841 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     6901 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     6961 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     7021 atgtt
//
