LOCUS       sequence-271736-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HAOccludinCT3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  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 6999)
  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-HAOccludinCT3
FEATURES             Location/Qualifiers
     source          1..6999
                     /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    1441..2063
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1640..1659)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2087..2107)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2105..2402
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2456..3250
                     /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(2510..2529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3120..3139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3902..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3939..3958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3993..4012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4041..4060)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4042..4060)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4074..4090)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4074..4090)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4087..4109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4098..4114
                     /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(4122..4128,4129..4146,4147..4152))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4167..4188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4305..4322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4476..5064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4556..4575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5235..6026,6027..6095))
                     /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     5858..5877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6096..6200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6227..6682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6314..6333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6524..6545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6808..6830
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6822..6839
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6823..6839
                     /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 acgctagcta 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 ggccaactgc agccaggcct acggcagcac ctacctcaac cagtacatct
     1321 accactactg caccgtggac ccccaggagg ctgtggctgc tgtctgtggg ttcctcatcg
     1381 tcatcctgct ctgcctcatc tgcttcttcg cccagaagac gcgctaatct agaggatccc
     1441 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     1501 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     1561 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     1621 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     1681 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     1741 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     1801 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     1861 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     1921 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     1981 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     2041 actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag
     2101 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2161 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2221 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2281 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2341 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     2401 ggaggaattg atcagcttgg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     2461 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2521 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2581 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2641 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2701 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2761 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2821 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2881 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2941 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3001 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3061 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3121 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3181 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3241 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3301 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3361 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3421 cccgggctcg atcccctcgc gagttggttc agctgctgcc tgaggctgga cgacctcgcg
     3481 gagttctacc ggcagtgcaa atccgtcggc atccaggaaa ccagcagcgg ctatccgcgc
     3541 atccatgccc ccgaactgca ggagtgggga ggcacgatgg ccgctttggt cgacccggac
     3601 gggacgctcc tgcgcctgat acagaacgaa ttgcttgcag gcatctcatg agtgtgtctt
     3661 cccgttttcc gcctgaggtc actgcgtgga tggagcgctg gcgcctgctg cgcgacggcg
     3721 agctgctcac cacccactcg ccaagctgga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgccgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3841 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3901 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3961 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4021 ttatcatgtc tggatcaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag
     4081 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4141 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4201 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4261 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5341 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6121 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6181 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     6241 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6301 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6361 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6421 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6481 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6541 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6601 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     6661 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     6721 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     6781 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     6841 ccaagctgat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     6901 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     6961 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
